Gene encoding IGG FC region-binding protein

ABSTRACT

A gene encoding an IgG Fc region-binding protein (FCgammaBP); a recombinant vector containing this gene; host cells transformed by this recombinant vector; a process for producing a recombinant protein which is obtained by incubating these host cells; and a protein having a recombinant IgG Fc region-binding activity which is obtained by the above-mentioned process.

SPECIFICATION

1. Technical Field

This invention relates to a gene encoding an IgG Fc portion-binding protein [hereinafter referred to as FcγBP (Fcγ Binding Protein) or IgG Fc BP), which is a protein specifically binding to the Fc portion of immunoglobulin G (IgG), a recombinant vector containing this gene, host cells transformed by this recombinant vector, a process for producing a recombinant protein which is obtained by incubating these host cells and a protein having a recombinant IgG Fc portion-binding activity which is obtained by the above-mentioned process.

2. Background Art

As one of immunocompetent cells, a macrophage incorporates a foreign invader or a complex thereof with immunoglobulin G (IgG) into the cell via phagocytosis and then digests the same. It is also capable of exerting an antigen presentation effect so as to induce the production of an antibody by lymphocytes. The main entry of this phagocytosis is an Fc receptor [FcγR (Fcγ receptor)] of IgG located on the surface of the macrophage cell. The inherent function of this Fcγ receptor, which is a receptor participating in the binding of the Fc portion of IgG, is the elimination of a pathogen or an antigen-IgG immune complex coated with IgG (opposition).

It has been known that Fcγ receptors may be roughly classified into three types including RI, RII and RIII [J. V. Ravetch and J.-P. Kinet, Annu. Rev. Immunol. (1991), 9:457-492]. The cDNA of each of these receptors has been successfully cloned. From 1990 to 1991, furthermore, several groups of workers found out proteins which were associated with these Fcγ receptors and required for the initiation of the incorporation of the binding matters by Fcγ receptors, thus providing a clue to clarify the switch mechanism [L. L. Lanier, G. Yu and J. H. Phillips, Nature (1989), 342:803-805; T. Kurosaki and J. V. Ravetch, Nature (1989), 342:805-807; D. G. Orioff, C. Ra, S. J. Frank, R. D. Klausner and J.-P. Kinet, Nature (1989), 347:189-191; P. Anderson, M. Caligiuri, C. O'Brien, T. Manley, J. Ritz and S. F. Schlossman. Prc. Natl. Acad. Sci. USA (1990), 87:2274-2278; T. Kurosaki, I. Gander and J. V. Ravetch, Proc. Natl. Acad. Sci. USA (1991), 88:3837-3841; L. Azzoni, M. Kamoun. T. W. Salcedo, P. Kanakaraj and B. Perussia, J. Exp. Med. (1992), 176:1745-1750; A. T. Ting, L. M. Karnitz, R. A. Schoon, R. T. Abraham and P. J. Leibson, J. Exp. Med. (1992), 176:1751-1755].

On the other hand, Kobayashi et al., reported a protein FcγBP, which could bind specifically to the Fc portion of IgG and was different from the conventionally known Fcγ receptors, occurring in human small intestinal and colonic epithelial cells, in particular, goblet cells. The binding of this protein specific to the IgG Fc portion was confirmed by using horse radish peroxidase-labeled matters. Namely, FcγBP bound not to IgGFab, IgA or IgM but exclusively to the Fc portion of IgG. Also, FcγBP underwent no cross reaction with antibodies of Fcγ receptors I, II and III [K. Kobayashi, M. J. Blaser and W. R. Brown, J. immunol. (1989), 143:2567-2574].

Further, they partially purified FcγBP from human colonic epithelial cells and constructed mouse monoclonal antibodies with the use of this protein as an antigen. Thus it was confirmed that FcγBP bound not only to these antibodies but also to mouse IgG [K. Kobayashi, Y. Hamada, M. J. Blaser and W. R. Brown, J. Immunol. (1991), 146:68-74].

The partially purified FcγBP was SDS (sodium dodecyl sulfate)-electrophoresed and then subjected to Western blotting with the use of the monoclonal antibodies. As a result, it was found out that FcγBP formed an associate of 200 kDa or above containing a protein of 78 kDa [K. Kobayashi, Y. Hamada, M. J. Blaser and W. R. Brown, J. Immunol. (1991), 146:68-74].

The above-mentioned FcγBP is identical with the Fcγ receptors in the point of being capable of binding to the Fc portion of IgG. However, the stability and structure of FcγBP as a protein and its role in vivo still remained unknown. It is, therefore, highly interesting to analyze FcγBP so as to clarify its structure and function.

As will be described hereinafter, it is assumed that FcγBP is secreted onto mucosae together with mucus and traps pathogens or viruses invading the body into the mucus to thereby facilitate the excretion of these invaders, thus participating in the mechanism of protection against infection. An autoantibody produced in excess in a mucosa suffering from inflammation activates the complement system or causes cytotoxicity by macrophages, etc., thus worsening the inflammation. It is assumed that FcγBP blocks the Fc portion of such an autoantibody to thereby inhibit the progression of the inflammation. Because of having these functions, FcγBP might be applicable to drugs, for example, agents for protecting infection, antiinflammatory (antiphlogistic) agents or diagnostic drugs for autoimmune diseases such as ulcerative colitis and Crohn's disease, etc.

To employ FcγBP for these medicinal purposes, it is required to obtain FcγBP in a large amount and in a uniform state. However, FcγBP in a uniform state can be hardly obtained in a large amount by the method wherein FcγBP is isolated from an animal tissue per se or a culture supernatant of FcγBP-producing cells. It is, therefore, required to produce FcγBP in a large amount by using gene recombination techniques.

The present inventors successfully identified the base sequence of a gene encoding FcγBP by cloning the cDNA of FcγBP with the use of monoclonal antibodies against FcγBP.

This cDNA was inserted into an appropriate vector and host cells were transformed by the expression vector thus obtained. Then the obtained transformant was incubated and the target protein thus produced was separated and purified. As a result, it was found out that the protein thus produced had a characteristic of binding specifically to human IgG. Thus, the present inventors have clarified that FcγBP can be produced in a large amount and in a uniform state.

DISCLOSURE OF THE INVENTION

Accordingly, the present invention provides a gene encoding FcγBP.

The present invention further provides a recombinant vector containing a gene encoding FcγBP.

The present invention furthermore provides procaryotic or eucaryotic host cells transformed by a recombinant vector containing a gene encoding FcγBP.

The present invention furthermore provides a process for producing FcγBP which comprises incubating host cells transformed by a recombinant vector containing a gene encoding FcγBP and separating and purifying the protein thus produced.

The present invention furthermore provides a protein showing an IgG Fc region-binding activity which is produced by the above-mentioned process.

The present invention furthermore provides a method for using a gene encoding FcγBP or a part of the same as a probe in Northern blotting or in situ hybridization for the identification of the tissue synthesizing FcγBP mRNA.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an electrophorogram showing the result of the hybridization of FcγBP mRNA performed with the use of the probe Q.

FIG. 2 shows the relations among the partial cDNA (about 7.8 kbp) employed in the expression of FcγBP and the clones NZ4, C72, Y1, X1 and V11.

FIGS. 3A-3C provide morphological photographs proving the expression of the protein in COS7 cells transformed by using the vector pNV11-SR. As the primary antibody, the K9 monoclonal antibody-producing hybridoma culture supernatant was used in 3A, while the K17 monoclonal antibody-producing hybridoma culture supernatant was used in 3B. In 3C, no primary antibody was added (i.e., the control case). In each case, the HRP-labeled goat antimouse IgG (H+L) F(ab′)₂ fragment was added as the secondary antibody.

FIGS. 4A-4C provide morphological photographs showing the ability of the protein, which had been expressed in COS7 cells transformed by using the vector pNV11-SR, to bind to human IgG. In each case, HRP-labeled human IgG was employed as the primary antibody. The secondary antibodies employed as the antagonist are as follows; 4A: none (i.e., the control case), 4B: a human IgG fraction purified by chromatography, and 4C: a human IgG Fc fraction purified by chromatography.

FIGS. 5A-5D provide morphological photographs showing the ability of the protein, which had been expressed in COS7 cells transformed by using the vector pNV11-SR, to bind to human IgG. In each case, HRP-labeled human IgG was employed as the primary antibody. The secondary antibodies employed as the antagonist are as follows; 5A: a human IgG F(ab′)₂ fraction purified by chromatography, 5B: a human IgM fraction purified by chromatography, 5C: human serum IgA purified by chromatography, and 5D: human secretor IgA purified by chromatography.

FIG. 6 is an electrophorogram of Northern blotting which shows the specificity of the expression of FcγBP mRNA in human tissues.

FIG. 7 is an electrophorogram showing the Northern blotting analysis of FcγBP mRNA in colonic mucosal epithelial cells wherein use was made of the probes Q, A and Y.

FIG. 8 shows the structure of the full length cDNA of FcγBP.

FIGS. 9A-9B provide morphological photographs showing the abilities of COS7 cells, which had been transformed by using the plasmids piF-A53 and piF-A8, to bind to a mixture of monoclonal antibodies K9/K17. 9A: piF-A53, and 9B: piF-A8.

FIG. 10 shows a comparison between the base sequences of exon/intron boundary region of FcγBP. The boxed capital letters show exons, while the small letters show introns. The underlined parts show highly conserved sequences. R: purine, and y: pyrimidine.

FIG. 11 shows the base sequence in the neighborhood of the initiation site on the 5′ side of FcγBP genomic DNA wherein capital letters show exons while small letters show introns. The underlined parts are identical with the sequence of the cDNA. The shaded thick letters show the TGA stop codon in frame and the ATG codon which is assumed to be the initiation site. Bases in the exons are exclusively numbered by referring the 5′ end of the cDNA clone NZ4 as to +1.

FIG. 12 shows the structure of human FcγBP and the locations therein corresponding to the clones employed in the present invention.

FIGS. 13A-13B provide morphological photographs showing CHO cells expressing the FcγBP fragment therein. The sample (13A) had been treated with 6.4 μM of methotrexate but no sodium butyrate, while the sample (13B) had been treated with 6.4 μM of methotrexate and 5 mM of sodium butyrate.

DETAILED DESCRIPTION OF THE INVENTION

The cDNA encoding FcγBP can be obtained by, for example, preparing the mRNA from cells producing FcγBP and converting the mRNA into the double stranded cDNA by a known method. Although human colonic mucosal epithelial cells are employed in the present invention as the mRNA source, use may be made of homogenates of tissues in which FcγBP is distributed (for example, human small intestine, duodenum, stomach, submandibular gland, sublingual gland, common bile duct, bronchus, etc.) without restriction. Also, it is known that a cell line HT29-18-N2 originating in human colonic cancer cells and its subspecies produce FcγBP. Thus, use may be made of these cell lines as the mRNA source.

In the present invention, a modified AGPC method (P. Chomczynski et al., Analytical Biochem., 162:156-159, 1987) is employed to prepare the mRNA. Alternatively, all RNAs may be prepared in accordance with the method of Chirgwin et al. (Biochemistry 18:5294-5299, 1979). It is also possible to prepare the mRNA by the methods employed in cloning the genes of other physiologically active proteins, for example, treating with a surfactant in the presence of a ribonuclease inhibitor (for example, a vanadium complex) or treating with phenol.

To obtain the double stranded DNA from the mRNA thus obtained, reverse transcription is performed by using the mRNA as a template and an oligo(dT) or random primer complementary to the polyA-chain at the 3′ end or a synthetic oligonucleotide corresponding to a part of the amino acid sequence of FcγBP as a primer to thereby synthesize a DNA complementary to the mRNA (i.e., the cDNA).

In the present invention, the cDNA is constructed by the reverse transcription from the polyadenylated RNA in accordance with the modified Gubler & Hoffman method with the use of a cDNA synthesis kit manufactured by Amersham or InVitrogen and random primers.

Then an adaptor is ligated to this cDNA followed by the insertion into the EcoRI site of the λgt11 vector. The cDNA library thus constructed is packaged into the λ phage by using an in vitro packaging kit Gigapack II Gold manufactured by Stratagene and expressed in Escherichia coli. The cDNA protein thus expressed is screened with the use of monoclonal antibodies as a probe.

As the antibodies to be used in the above-mentioned cloning, those capable of detecting FcγBP in Western blotting are selected from among 3 monoclonal antibodies (K9, K10 and K17) which can inhibit the binding of IgGFc to FcγBP (Kobayashi et al., J. Immunology, 146:68-74, 1991; Kobayashi et al., J. Immunology, 143:2567-2574, 1989). Namely, when the antibody K9 is used, a band larger than about 200 kDa is observed under reducing conditions. When the antibody K17 is used, on the other hand, bands at 70-80 kDa and 130-140 kDa are observed under nonreducing conditions. Based on these results, these monoclonal antibodies capable of recognizing 2 different epitopes are employed in the cloning.

As the result of this screening, one clone, which is named probe Q (600 bp), is obtained from about 1,000,000 clones by using the antibody K9, while 7 clones, among which a DNA fragment of 700 bp is named probe A and another DNA fragment of 600 bp is named probe B, are obtained from about 600,000 clones by using the antibody K17.

By using this probe Q, the size of the FcγBP mRNA is estimated through comparison with the mRNAs of known proteins. As a result, it is estimated that the FcγBP mRNA molecule has a size of about 17 kbp (FIG. 1).

Next, the second cDNA library packaged into λgt10 is screened with the use of the probes Q, A and B. First, cDNA clones hybridizable with one of the probes Q, A and B are separated. From among these clones, one hybridizable exclusively with the probe A is obtained. The end of this clone on the opposite side to A is referred to as probe X (about 700 bp) and a cDNA clone hybridizable with this probe X is separated. The obtained clone, which has X at the center and the A-B region at one end, is named X1. Subsequently, a part (about 800 bp) of this clone X1 opposite to the A-B region is named probe Y and the cDNA library is screened again. Thus a cDNA clone hybridizable with this probe Y is obtained and named Y1. This clone Y1 has a part of the X region at one end and the Y region at the center. A part (about 150 bp) of this clone Y1 at the opposite end to the X region is named probe Y150. By using this probe Y150, the cDNA library is screened again. Thus a clone showing the longest extension on the opposite side to the Y150 region is selected and named clone C72. Subsequently, a part (about 450 bp) of this clone C72 in the neighborhood of the opposite end to the Y150 region is named probe Z and cDNA clones hybridizable with this probe Z are separated. From among these clones, one having the longest part not overlapping the clone C72 is separated and named clone NZ4.

From among the cDNA clones hybridizable each of the clones A, B and Q, on the other hand, one having the same base sequence in the A-B region as that of the A-B region in the clone X1 is selected and named clone V11.

Then the base sequences of these 5 clones (X1, Y1, C72, NZ4 and V11) are identified and the amino acid sequences of the proteins are confirmed. Thus an open reading frame having ATG as the initiation codon is found out (FIG. 2).

Further, a partial cDNA (about 7.8 kbp) encoding FcγBP is obtained from these clones and its base sequence is identified (SEQ ID NO:6 shows the base sequence and the amino acid sequence).

Furthermore, the cDNA clones obtained by screening via the hybridization with the use of the above-mentioned probe A, B or Q are each amplified in Escherichia coli and mapped by using the probes A, B and Q. As a result, it is assumed that the cDNA of FcγBP has a structure of 16.4 kbp in the full length with a unit of 3.5 kbp, wherein sequences homologous respectively with the probes A, B and Q are linked together in the order of A→B→Q, repeated in tandem.

In the cDNA clone hybridizable with the probe B, a part of the base sequence of the probe is amplified by PCR and the base sequence of the fragment thus obtained is analyzed. Thus it is clarified that the full-length cDNA of FcγBP has the structure shown in FIG. 8 and the base sequence and amino acid sequence represented by SEQ ID NO:7 in Sequence Listing.

By integrating the gene encoding FcγBP thus cloned into an appropriate vector, procaryotic or eucaryotic host cells can be transformed.

Furthermore, an appropriate promoter or a sequence relating to the phenotypic expression may be introduced into such a vector to thereby express the gene in host cells. It is also possible that the target gene is ligated with an additional gene encoding another polypeptide and thus expressed as a fused protein so as to facilitate the purification or elevate the expression yield. Also, the target protein can be excised by performing an appropriate treatment in the process of the purification.

It is generally considered that a eucaryotic gene shows polymorphism as is the case with that for human interferon. Owing to this polymorphism, therefore, one or more amino acids are replaced in some cases, or every amino acid remains unchanged regardless of changes in the base sequence in other cases.

Accordingly, it is sometimes observed that a polypeptide which has the deletion, addition or replacement of one or more amino acids in the amino acid sequence represented by SEQ ID NO:6 or SEQ ID NO:8 in Sequence Listing or a part of the same has the IgG Fc portion-binding activity. For example, it is publicly known that when the base sequence corresponding to cysteine in human interleukin 2 (IL-2) gene is converted into another base sequence corresponding to serine, the polypeptide thus obtained still sustains the IL-2 activity (Wang et al., Science, 224:1431, 1984).

When polypeptides are expressed in eucaryotic cells, glycosylation occurs in many cases. The glycosylation may be regulated by converting one or more amino acids. In such a case, the obtained polypeptide sometimes has the IgG Fc portion-binding activity. When the FcγBP gene of the present invention is artificially modified and the polypeptides thus obtained have the IgG Fc portion-binding activity, therefore, genes encoding these polypeptides are all involved in the scope of the present invention.

The present invention further involves genes encoding polypeptides which have the IgG Fc portion-binding activity and are hybridizable with the genes represented by SEQ ID NO:6 or SEQ ID NO:7 in Sequence Listing or a part of the same. The hybridization may be carried out under the conditions commonly employed in probe hybridization (refer to, for example, Molecular Cloning: A Laboratory Manual, Sambrook et al., Cold Spring Harbor Laboratory Press, 1989).

As described above, it is clear that the present invention involves not only the base sequences encoding the amino acid sequences represented by SEQ ID NO:6 and SEQ ID NO:7 in Sequence Listing but also variously modified DNAs so long as they are capable of expressing proteins binding specifically to the Fc portion of immunoglobulin (IgG). Also, DNA fragments having the above-mentioned function fall within the scope of the present invention.

The expression vector of the present invention involves a replication source, a selective marker, a promoter, an RNA splice site, a polyadenylation signal, etc.

Examples of the procaryotic host cells to be used in the expression system of the present invention include Escherichia coli, Bacillus subtilis, etc. Examples of eucaryotic host cells usable herein include yeasts and slime molds. Also, use can be made of insect cells such as Sf9 as the host cells. Further, examples of host cells originating in animal cells include COS, CCHO, C127 and 3T3 cells.

Thus the host cells are transformed by the gene encoding the target FcγBP and the resulting transformant is incubated. Then the protein having the IgG Fc region-binding activity thus produced can be intracellularly or extracellularly separated and purified.

The methods for the separation and purification of the protein having the IgG Fc portion-binding activity, namely, the target protein of the present invention are not restricted to those employed in the Examples given hereinafter but can be arbitrarily selected from among the methods commonly employed in the separation and purification of proteins. For example, various chromatographic procedures, ultrafiltration, salting out, dialysis, etc. may be appropriately selected and combined therefor.

Then it is examined whether or not the recombinant protein thus obtained has the activity of binding to human IgG similar to the native FcγBP. As a result, it is proved that this recombinant protein binds specifically to human IgGFc.

As described above, the full-length cDNA of FcγBP of the present invention has the structure shown in FIG. 8 and the base sequence and the amino acid sequence represented by SEQ ID NO:7 in Sequence Listing. Further, it is confirmed again in the following manner that a series of cDNAs employed in the present invention originate in a single mRNA. That is, cDNA fragments in a repeating structure different from pNV11SR including the 5′ end cDNA employed in the expression are expressed and reacted with the monoclonal antibodies K9 and K17 which recognize FcγBP. As a result, it is confirmed that these cDNA products are recognized by either or both of K9 and K17.

Further, the tissue-specificity of the expression of the mRNA of FcγBP is examined. Thus it is found out that this protein is expressed in human placenta. Accordingly, it is possible to identify a tissue synthesizing the FcγBP mRNA by Northern blotting or in situ hybridization with the use of the gene encoding FcγBP of the present invention or a part of the same as a probe.

Furthermore, the occurrence of polymorphism on the chromogen of FcγBP can be proved by using restriction enzymes.

The following Examples will be given in order to illustrate a process for obtaining the gene encoding FcγBP of the present invention, a recombinant vector containing this gene, a transformant containing this vector, the target protein obtained by incubating this transformant and processes for producing each of them. However, it is to be understood that the present invention is not restricted thereto.

EXAMPLES Example 1 Cloning of Partial cDNA Encoding FcγBP by Using Monoclonal Antibody (A: Construction of cDNA library)

(1) Preparation of Human Colonic Mucosal Epithelial Cell

A human colonic tissue piece was thoroughly washed with RPMI medium containing 10% of FBS and mechanically peeled off from the muscularis mucosae to thereby separate the epithelial cells from the lamina propria mucosae. Next, it was fixed to the shaft and vigorously stirred with a stirrer in 10% FBS/5 mM EDTA/PBS (−) for 90 minutes on ice to thereby separate the epithelial cells. Then the solution containing the epithelial cells was centrifuged at 1,500 rpm for 10 minutes to thereby give a precipitate of the cells.

(2) Purification of mRNA

All RNAs were extracted from the mucosal epithelial cells by a modified AGPC method (P. Chomczynski et al., Analytical Biochem., 162:156-159, 1987). Namely, to 1 ml of the cell pellets was added 9 ml of a denaturation solution [4 M of guanidine thiocyanate, 25 mM of sodium citrate (pH 7), 0.5% of Sarkosyl, 0.1 M of 2-mercaptoethanol]. After lysing the cells, 1 ml of 2 M sodium acetate (pH 4), 10 ml of a saturated aqueous solution of phenol and 2 ml of chloroform/isoamyl alcohol (49:1) were successively added thereto. Then the mixture was stirred for 10 seconds, ice-cooled for 15 minutes and then centrifuged at 10,000×g for 15 minutes and the supernatant was taken up. To 8 ml of this supernatant were similarly added 0.8 ml of sodium acetate, 8 ml of water-saturated phenol and 1.6 ml of chloroform/isoamyl alcohol. Then the mixture was stirred for 10 seconds, ice-cooled for 15 minutes and centrifuged at 10,000×g for 15 minutes and the supernatant was taken up. To 7 ml of this supernatant was added the same amount of chloroform/isoamyl alcohol and the obtained mixture was stirred and centrifuged to thereby give the supernatant. To the obtained supernatant was added the same amount of isopropanol and the mixture was cooled at −20° C. for 30 minutes and then centrifuged at 10,000×g for 15 minutes to thereby recover the precipitate of all RNAs.

To a solution of 1 mg of the all RNAs, an elution buffer [10 mM of Tris-HCl (pH 7.5), 1 mM of EDTA, 0.1% of SDS] was added so as to adjust the total volume to 1 ml. Then 1 ml of OligoTex-dT30 <Super> (manufactured by Takara Shuzo) was added thereto and the mixture was heated at 65° C. for 5 minutes and quenched on ice for 3 minutes. After adding 0.2 ml of 5 M NaCl and maintained at 37° C. for 10 minutes, the mixture was centrifuged at 15,000 rpm for 3 minutes and the supernatant was carefully removed. The pellets were suspended in 2.5 ml of a washing buffer [10 mM of Tris-HCl (pH 7.5), 1 mM of EDTA, 0.5 M of NaCl, 0.1% of SDS] and centrifuged at 15,000 rpm for 3 minutes and the supernatant was carefully removed. The pellets were suspended in 1 ml of sterilized water, heated at 65° C. for 5 minutes and then quenched on ice for 3 minutes. Then it was centrifuged at 15,000 rpm for 3 minutes and the supernatant was taken up. To the supernatant were added 50 μl of 5 M NaCl and 2.5 ml of ethanol and the mixture was cooled at −20° C. for 30 minutes and centrifuged (3,000 rpm, 4° C.). Then the precipitate of the polyadenylated RNA was recovered.

(3) Synthesis of cDNA

From the mRNA, cDNA was synthesized by a modified method of Gubler and Hoffman [U. Gubler and B. J. Hoffman (1983), Gene, 25:263] with the use of a cDNA synthesis kit manufactured by Amersham or InVitrogen. Namely, 5 μg of the polyadenylated RNA prepared from the colonic mucosal epithelial cell was incubated at 42° C. for 90 minutes in 50 μl of a buffer (manufactured by Amersham) containing 50 U of human placenta ribonuclease inhibitor, 1 mM of dATP, 1 mM of dGTP, 1 mM of dCTP, 0.5 mM of dTTP, 100 U of AMV reverse transcriptase and sodium pyrophosphate together with 750 ng of random hexanucleotide or 4 μg of oligo(dT) primer. 50 μl of this reaction mixture was reacted in a buffer (manufactured by Amersham) containing 4.0 U of Escherichia coli ribonuclease H and 115 U of Escherichia coli DNA polymerase I successively at 12° C. for 60 minutes and at 22° C. for 60 minutes and then incubated at 70° C. for 10 minutes. After returning into ice, 10 U of T4 DNA polymerase was added thereto and reacted at 37° C. for 10 minutes. Next, the reaction was ceased by adding 10 μl of 0.25 M EDTA (pH 8). To 250 μl of the reaction mixture were added the same amount of 7.5 M ammonium acetate and 4 times as much ethanol. After stirring, the mixture was cooled at −20° C. for 30 minutes and centrifuged to thereby collect cDNA. The cDNA was dissolved in 10 μl of sterilized water and 1 μl of this solution was electrophoresed on a 0.8% agarose gel to confirm the synthesis and determine the concentration.

(4) Ligation of Adaptor

To the cDNA synthesized in the above (3) was added 10 times by mol as much an adaptor (EcoRI-NotI-BamHI adaptor, manufactured by Takara Shuzo). Then a ligation solution A (ligation kit, manufactured by Takara Shuzo) in an amount 8 times more than the total mixture and a ligation solution B (ligation kit, manufactured by Takara Shuzo) in the same amount as the total mixture were added thereto. After thoroughly stirring, the resulting mixture was incubated at 16° C. for 30 minutes to thereby ligate the adaptor to the cDNA.

This reaction mixture was electrophoresed on a 1% low melting agarose gel (Sea Plaque Agarose, manufactured by Takara Shuzo) in a TAE buffer system and thus a gel containing a cDNA fraction of 0.5 kbp or above was recovered. By this procedure, the adaptor not ligated to the cDNA was eliminated at the same time. Then TE buffer in an amount twice more than the wet weight of the gel recovered above was added thereto and the mixture was maintained at 65° C. for 10 minutes. After thus dissolving the agarose gel, Tris-saturated phenol in the same amount as the total mixture was added thereto. Then the resulting mixture was thoroughly stirred, ice-cooled and centrifuged. The aqueous phase was taken up and treated with the same amount of Tris-saturated phenol again. After centrifuging, the aqueous phase was taken up and the same amount of chloroform was added thereto. The mixture was thoroughly stirred and centrifuged. Next, the aqueous phase was taken up and 3 M sodium acetate in an amount {fraction (1/10)} times as much, 20 μg of glycogen (manufactured by Boehringer-Mannheim) and ethanol in an amount 2.5 times as much were added thereto. After cooling at −20° C. for 30 minutes, the mixture was centrifuged at 15,000 rpm for 10 minutes at 4° C. to thereby give a precipitate of the cDNA.

(5) Construction of λgt11 Library

The cDNA obtained in the above (4), to which the adaptor had been ligated, was dissolved in 96 μl of a solution comprising 500 mM of Tris-HCl (pH 7.5), 100 mM of MgCl₂, 10 mM of DTT and 10 mM of ATP. After adding 40 U of polynucleotide kinase, the mixture was incubated at 37° C. for 60 minutes to thereby phosphorylate the 5′ end of the adaptor. After the completion of the reaction, 200 μl of TE buffer and 300 μl of Tris-saturated phenol were successively added thereto. The obtained mixture was stirred and centrifuged (15,000 rpm, room temperature, 2 minutes) and the supernatant was taken up. Similarly, centrifugation was performed by successively using a Tris-saturated phenol/chloroform (1:1) solution and a chloroform solution containing 2% of isoamyl alcohol to thereby give 250 μl of the supernatant finally. To this supernatant were added 250 μl of a 4 M ammonium acetate solution and 1,250 μl of ethanol. The obtained mixture was cooled at −20° C. for 30 minutes and then centrifuged (15,000 rpm, 4° C., 10 minutes) to thereby recover the precipitate. To the cDNA precipitate was added 1 μg of EcoRI-digested dephosphorylated λgt11 arm (#234211, manufactured by Stratagene) and dissolved in 5 μl of a solution containing 100 mM (the final concentration) of Tris-HCl (pH 7.6), 5 mM of MgCl₂ and 300 mM of NaCl. Then 5 μl of the ligation solution B (DNA ligation kit, manufactured by Takara Shuzo) was added and the mixture was thoroughly stirred and then reacted at 26° C. for 10 minutes. To package the cDNA into the λ phage, 4 μl of the ligation mixture containing the cDNA was added to 10 μl of Freeze/Thaw extract (Gigapack II gold, manufactured by Stratagene). Immediately thereafter, Sonic extract (Gigapack II gold) was added thereto and the resulting mixture was slowly stirred. After incubating at 22° C. for 2 hours, 500 μl of a solution for diluting the phage [5 g of NaCl, 2 g of MgSO₄.7H₂O, 50 ml of 1M Tris-HCl (pH 7.5), 5 ml of 2% gelatin per liter] and 10 μl of chloroform were added and thus the in vitro packaging reaction was completed. This phage solution was stored at 4° C. and employed in screening.

Example 2 Cloning of Partial cDNA Encoding FcγBP by Using Monoclonal Antibody (B: Screening of cDNA library by using antibody)

(1) Screening

1×10⁴ pfu (200 μl) of the cDNA library of the colonic mucosal epithelial cells packaged into the λ phage, which had been prepared in the above Example 1, was mixed with 200 μl of an Escherichia coli strain Y1090γ⁻, which had been incubated overnight, and then incubated at 37° C. for 15 minutes. Separately, 0.8% top agarose gel was mixed with LB medium, dissolved therein and maintained at 55° C. Then 5 ml of this material was added to the preincubation of the phage with Escherichia coli and mixed homogeneously. The obtained mixture was uniformly spread onto a 1.5% LB agarose plate (10×14 cm) and stored in an incubator at 42° C. for 3.5 hours. After confirming that small plaques had been formed, a nylon-reinforced nitrocellulose filter (#BA-S85, manufactured by Schleicher & Schnell), which had been impregnated with IPTG and air-dried, was placed on the plate followed by storage at 37° C. for 3.5 hours. Then the filter was peeled off from the plate and shaken in a washing liquor [0.05% of Tween-20, 25 mM of Tris-HCl (pH 7.5), 150 mM of NaCl, 3 mM of KCl] at room temperature for 30 minutes. Subsequently, it was shaken in PBS (−) containing 5% of skim milk at room temperature for 30 minutes for blocking and then washed with the washing liquor for 20 minutes twice. Then the nitrocellulose membrane was immersed in 5 ml of a hybridoma culture supernatant containing the mouse monoclonal antibody K9 or K17 which had been prepared against FcγBP in colonic mucosal epithelial cells [Kobayashi et al., J. Immunology (1991), 146:68-74; Kobayashi et al., J. Immunology (1989), 143:2567-2574]. After shaking at room temperature for 2 hours, the membrane was washed with the washing liquor for 20 minutes twice. Next, it was shaken in horse radish peroxidase (HRP)-labeled antimouse IgG (H+L) goat antiserum (manufactured by Zymed), which had been diluted 1,000-fold with the washing liquor, at room temperature for 1 hour and washed with the washing liquor for 20 minutes twice. After washing with a TBS solution free from Tween-20 for 10 minutes, it was immersed in a mixture of 50 ml of a diaminobenzidine solution [1 mg/ml of 0.1 M Tris-HCl (pH 7.2)], 50 ml of a 0.02% solution of H₂O₂ and 50 μl of an 8% solution of NiCl₂ to thereby detect positive plaques.

(2) Extraction of λDNA

By the screening with the use of the monoclonal antibody K9, a clone containing a cDNA insert of about 600 bp was obtained from among about 1,000,000 plaques. This plaque was picked up with a toothpick and the λ phage was incubated in 200 μl of a medium (LB medium containing 20 mM of MgSO₄, 0.2% of maltose and 5 μl of an overnight culture suspension of Escherichia coli Y1090γ⁻ strain) at 37° C. for 4 hours. 2 μl (1×10⁷ pfu/μl) of the culture medium containing the phage was added to 10 ml of LB medium (containing 20 mM of MgSO₄ and 0.25 ml of an overnight culture suspension of Escherichia coli Y1090γ⁻ strain) which was thus infected therewith. Then the λ phage was propagated by incubating at 37° C. for 5 hours under shaking.

After incubating for 5 to 6 hours and confirming the occurrence of bacteriolysis, 50 μl of chloroform and 2 ml of 5 M NaCl were added followed by shaking at 37° C. for 10 minutes. Then the mixture was centrifuged at 3,500 rpm for 15 minutes. To the supernatant thus obtained was added polyethylene glycol 6000 in such an amount as to give a concentration of 10%. The resulting mixture was allowed to stand on ice for 30 to 60 minutes and then centrifuged at 4,000 rpm for 15 minutes at 4° C. to thereby precipitate the phage. The precipitate was suspended in 1 ml of A buffer [0.5% of NP-40, 30 mM of Tris-HCl (pH 7.5), 5 mM of MgCl₂, 125 mM of KCl, 3.6 mM of CaCl₂, 0.5 mM of EDTA, 0.25% of sodium deoxycholate, 60 mM of mercaptoethanol] and incubated together with 100 μg/ml of RNase A and 20 μg/ml of DNase I at 37° C. for 30 minutes. Then chloroform in the same amount of the A buffer was added thereto and the mixture was stirred and centrifuged at 15,000 rpm for 2 minutes at room temperature followed by the recovery of the supernatant. Then the same amount of chloroform was added again and the mixture was centrifuged in the same manner followed by the recovery of the supernatant. To the supernatant were added 50 mM of Tris-HCl (pH 8), 20 mM of EDTA, 0.5% of SDS and 100 μg/ml of protease K and the resulting mixture was incubated at 55° C. for 60 minutes. To purify the λDNA, treatments with phenol, phenol/chloroform and chloroform were carried out each in the conventional manner so as to inactivate the DNAse, protease, etc. Then {fraction (1/20)} times as much 5 M NaCl and 1 time as much isopropanol were added thereto. Thus a precipitate of the λDNA having the cDNA fragment inserted therein was obtained.

(3) Construction of Probe DNA

From the λDNA having the cDNA fragment purified in the above (2), the DNA insert was excised with the use of the BamHI restriction enzyme site and employed as a probe for screening the second cDNA library (this probe was named probe Q).

By the same method, DNA probes of 700 bp and 600 bp were excised with BamHI from the clone having the longest insert (about 1,300 bp) among the 7 λ clones obtained by using the monoclonal antibody K17. In these probes, the DNA fragment of 700 bp, which seemingly contained the cDNA encoding the epitope of the antibody K17, was referred to as probe A, while another fragment of 600 bp was referred to as probe B. These probes were employed in screening the second cDNA library.

(4) Northern Blotting

It was confirmed by Northern blotting that the probes A and Q obtained by screening with the use of the antibodies were hybridizable with the same mRNA.

15 μg of all RNAs extracted from colonic mucosal epithelial cells by the AGPC method were dissolved in 4.5 μl of sterilized water, then mixed with 2 μl of 5×MOPS buffer, 3.5 μl of formaldehyde and 10 μl of formamide and thermally denatured at 60° C. for 15 minutes. Next, the denatured product was electrophoresed on a 1% agarose gel in the presence of formaldehyde. After the completion of the electrophoresis, the RNAs were transferred onto a nylon membrane (Biodyne A, manufactured by Pall Corp.) by the capillary method overnight. After fixing the RNAs on the nylon membrane by UV-crosslinking, prehybridization was performed in 10 ml of a hybridization solution (5×SSPE, 5×Denhardt's solution, 50% of formamide, 0.5% of SDS, 100 μg/ml of thermally denatured salmon sperm DNA) at 42° C. for 8 hours.

Subsequently, the probes A and Q obtained by the antibody screening were each radioisotope-labeled with the use of α[³²P]dCTP by Megaprime Labeling Kit (manufactured by Amersham). 1×10⁸ dpm of each probe was added together with 5 ml of the hybridization solution to the nylon membrane which had been subjected to the prehybridization. After sealing, hybridization was carried out at 42° C. overnight. The nylon membrane was washed in a solution containing 0.2×SSC and 0.2% of SDS at 65° C. for 40 minutes thrice. Then the nylon membrane was dried and exposed to an X-ray film overnight.

Thus a band of about 17 kbp was detected by using each of the probes A and Q, which proved that these 2 probes were hybridizable with the same mRNA from the viewpoint of molecular weight.

Example 3 Second Cloning of cDNA Encoding FcγBP (A: Construction of cDNA library)

(1) Preparation of Human Colonic Mucosal Epithelial Cell

A human colonic tissue piece was thoroughly washed with RPMI medium containing 10% of FBS and mechanically peeled off from the muscularis mucosae to thereby separate the epithelial cells from the lamina propria mucosae. Next, it was fixed to the shaft and vigorously stirred with a stirrer in 10% FBS/5 mM EDTA/PBS (1) for 90 minutes on ice to thereby separate the epithelial cells. Then the solution containing the epithelial cells was centrifuged at 1,500 rpm for 10 minutes to thereby give a precipitate of the cells.

(2) Purification of mRNA

All RNAs were extracted from the mucosal epithelial cells by a modified AGPC method [P. Chomczynski et al., Analytical Biochem., (1987) 162:156-159]. Namely, to 1 ml of the cell pellets was added 9 ml of a denaturation solution [4 M of guanidine thiocyanate, 25 mM of sodium citrate (pH 7), 0.5% of Sarkosyl, 0.1 M of 2-mercaptoethanol]. After lysing the cells, 1 ml of 2 M sodium acetate (pH 4), 10 ml of a saturated aqueous solution of phenol and 2 ml of chloroform/isoamyl alcohol (49:1) were successively added thereto. Then the mixture was stirred for 10 seconds, ice-cooled for 15 minutes and then centrifuged at 10,000×g for 15 minutes and the supernatant was taken up. To 8 ml of this supernatant were similarly added 0.8 ml of sodium acetate, 8 ml of water-saturated phenol and 1.6 ml of chloroform/isoamyl alcohol. Then the mixture was stirred for 10 seconds, ice-cooled for 15 minutes and then centrifuged at 10,000×g for 15 minutes and the supernatant was taken up. To 7 ml of this supernatant was added the same amount of chloroform/isoamyl alcohol and the obtained mixture was stirred and centrifuged to thereby give the supernatant. To the obtained supernatant was added the same amount of isopropanol and the mixture was cooled at −20° C. for 30 minutes and then centrifuged at 10,000×g for 15 minutes to thereby recover the precipitate of all RNAs.

To a solution of 1 mg of the all RNAs, an elution buffer [10 mM of Tris-HCl (pH 7.5), 1 mM of EDTA, 0.1% of SDS] was added so as to adjust the total volume to 1 ml. Then 1 ml of OligoTex-dT30 <Super> (manufactured by Takara Shuzo) was added thereto and the mixture was heated at 65° C. for 5 minutes and quenched on ice for 3 minutes. After adding 0.2 ml of 5 M NaCl and maintained at 37° C. for 10 minutes, the mixture was centrifuged at 15,000 rpm for 3 minutes and the supernatant was carefully removed. The pellets were suspended in 2.5 ml of a washing buffer [10 mM of Tris-HCl (pH 7.5), 1 mM of EDTA, 0.5 M of NaCl, 0.1% of SDS] and centrifuged at 15,000 rpm for 3 minutes and the supernatant was carefully removed. The pellets were suspended in 1 ml of sterilized water, heated at 65° C. for 5 minutes and then quenched on ice for 3 minutes. Then it was centrifuged at 15,000 rpm for 3 minutes and the supernatant was taken up. To the supernatant were added 50 μl of 5 M NaCl and 2.5 ml of ethanol and the mixture was cooled at −20° C. for 30 minutes and centrifuged (3,000 rpm, 4° C.). Then the precipitate of the polyadenylated RNA was recovered.

(3) Synthesis of cDNA

From the mRNA, cDNA was synthesized by a modified method of Gubler and Hoffman with the use of a cDNA synthesis kit manufactured by Amersham or InVitrogen. Namely, 5 μg of the polyadenylated RNA prepared from the colonic mucosal epithelial cells was incubated at 42° C. for 90 minutes in 50 μl of a buffer (manufactured by Amersham) containing 50 U of human placenta ribonuclease inhibitor, 1 mM of dATP, 1 mM of dGTP, 1 mM of dCTP, 0.5 mM of dTTP, 100 U of AMV reverse transcriptase and sodium pyrophosphate together with 750 ng of random hexanucleotide or 4 μg of oligo(dT) primer. 50 μl of this reaction mixture was reacted in a buffer (manufactured by Amersham) containing 4.0 U of Escherichia coli ribonuclease H and 115 U of Escherichia coli DNA polymerase I successively at 12° C. for 60 minutes and at 22° C. for 60 minutes and then incubated at 70° C. for 10 minutes. After returning into ice, 10 U of T4 DNA polymerase was added thereto and reacted at 37° C. for 10 minutes. Next, the reaction was ceased by adding 10 μl of 0.25 M EDTA (pH 8). To 250 μl of the reaction mixture were added the same amount of 7.5 M ammonium acetate and 4 times as much ethanol. After stirring, the mixture was cooled at −20° C. for 30 minutes and centrifuged to thereby collect cDNA. The cDNA was dissolved in 10 μl of sterilized water and 1 μl of this solution was electrophoresed on a 0.8% agarose gel to confirm the synthesis and determine the concentration.

(4) Ligation of Adaptor

To the cDNA synthesized in the above (3) was added 10 times by mol as much an adaptor (EcoRI-NotI-BamHI adaptor, manufactured by Takara Shuzo). Then a ligation solution A (ligation kit, manufactured by Takara Shuzo) in an amount 8 times more than the total mixture and a ligation solution B (ligation kit, manufactured by Takara Shuzo) in the same amount as the total mixture were added thereto. After thoroughly stirring, the mixture was incubated at 16° C. for 30 minutes to thereby ligate the adaptor to the cDNA.

This reaction mixture was electrophoresed on a 0.8% low melting agarose gel (Sea Plaque agarose gel, manufactured by Takara Shuzo) in a TAE buffer system and thus a gel containing a cDNA fraction of about 4 kbp or above was recovered. By this procedure, the adaptor not ligated to the cDNA was eliminated at the same time. Then TE buffer in an amount twice more than the wet weight of the gel recovered above was added thereto and the mixture was maintained at 65° C. for 10 minutes. After thus dissolving the agarose gel, Tris-saturated phenol was added in the same amount as the total mixture. Then the resulting mixture was thoroughly stirred, ice-cooled and centrifuged. The aqueous phase was taken up and treated with the same amount of Tris-saturated phenol again. After centrifuging, the aqueous phase was taken up and the same amount of chloroform was added thereto. The mixture was thoroughly stirred and centrifuged. Next, the aqueous phase was taken up and {fraction (1/10)} times as much 3 M sodium acetate, 20 μg of glycogen (manufactured by Boehringer-Mannheim) and 2.5 times as much ethanol were added thereto. After cooling at −20° C. for 30 minutes, the mixture was centrifuged at 15,000 rpm for 10 minutes at 4° C. to thereby give a precipitate of the cDNA.

(5) Construction of λgt11 Library

The cDNA obtained in the above (4), to which the adaptor had been ligated, was dissolved in 96 μl of a solution comprising 500 mM of Tris-HCl (pH 7.5), 100 mM of MgCl₂, 10 mM of DTT and 10 mM of ATP. After adding 40 U of polynucleotide kinase, the mixture was incubated at 37° C. for 60 minutes to thereby phosphorylate the 5′ end of the adaptor. After the completion of the reaction, 200 μl of TE buffer and 300 μl of Tris-saturated phenol were successively added thereto. The obtained mixture was stirred and centrifuged (15,000 rpm, room temperature, 2 minutes) and the supernatant was taken up. Similarly, centrifugation was performed by successively using a Tris-saturated phenol/chloroform (1:1) solution and a chloroform solution containing 2% of isoamyl alcohol to thereby give 250 μl of the supernatant finally. To this supernatant were added 250 μl of a 4 M ammonium acetate solution and 1,250 μl of ethanol. The obtained mixture was cooled at −20° C. for 30 minutes and centrifuged (15,000 rpm, 4° C., 10 minutes) to thereby recover the precipitate. To the cDNA precipitate was added 1 μg of EcoRI-digested dephosphorylated λgt10 arm (#233211, manufactured by Stratagene) and dissolved in 5 μl of a solution containing 100 mM (the final concentration) of Tris-HCl (pH 7.6), 5 mM of MgCl₂ and 300 mM of NaCl. Then 5 μl of the ligation solution B (DNA ligation kit, manufactured by Takara Shuzo) was added and the mixture was thoroughly stirred and then reacted at 26° C. for 10 minutes. To package the cDNA into the λ phage, 4 μl of the ligation mixture containing the cDNA was added to 10 μl of Freeze/Thaw extract (Gigapack II gold, manufactured by Stratagene). Immediately thereafter, Sonic extract (Gigapack II gold) was added thereto and the resulting mixture was slowly stirred. After incubating at 22° C. for 2 hours, 500 μl of a solution for diluting the phage [5 g of NaCl, 2 g of MgSO₄.7H₂O, 50 ml of 1M Tris-HCl (pH 7.5), 5 ml of 2% gelatin per liter] and 10 μl of chloroform were added and thus the in vitro packaging reaction was completed. This phage solution was stored at 4° C. and employed in screening.

Example 4 Cloning of Full Length cDNA Encoding FcγBP (B: Screening of cDNA library by using DNA probe)

(1) Blotting

200 μl of an Escherichia coli strain c600hfl, which had been incubated overnight, was infected with the colonic mucosal epithelial cell cDNA (2×10⁴ pfu) packaged into λ phage and then maintained at 37° C. for 15 minutes. After adding a 0.8% top agarose/LB medium maintained at 55° C., the mixture was immediately spread onto an LB plate (10×14 cm) and incubated at 37° C. for 12 hours. When the diameter of a plaque became about 1 mm, a nylon membrane (Biodyne A, pore size: 0.2μ, manufactured by Pall Corp.) was placed thereon followed by cooling at 4° C. for 10 minutes. Then the nylon membrane was peeled off from the plate and treated with the blotting solution I (0.5 M of NaOH, 1.5 M of NaCl), the blotting solution II [1 M of Tris-HCl (pH 7.4)] and the blotting solution III [0.5 M of Tris-HCl (pH 7.4), 1.5 M of NaCl] each for 5 minutes. Then the DNA was fixed onto the nylon membrane with the use of an UV-crosslinking device (UV Stratalinker 2400, manufactured by Stratagene) at 1,200 μJ.

(2) Hybridization

To a nylon membrane having the λDNA fixed thereon was added 10 ml of a hybridization solution (5×SSPE, 5×Denhardt's solution, 50% of formamide, 0.5% of SDS, 100 μg/ml of thermally denatured salmon sperm DNA). Next, the mixture was sealed in a hybridization bag and subjected to prehybridization at 42° C. for 8 hours. Subsequently, the probes Q, A and B obtained by the antibody screening were each radioisotope-labeled with the use of α[³²P]dCTP by the random priming method. 1×10⁸ dpm of each probe was added together with 5 ml of the hybridization solution to the nylon membrane which had been subjected to the prehybridization. After sealing, hybridization was carried out at 42° C. overnight. When the hybridization was completed, the nylon membrane was washed in a solution containing 0.2×SSC and 0.2% of SDS at 65° C. for 40 minutes thrice. Then the nylon membrane was dried and exposed to an X-ray film overnight.

By the above-mentioned screening, 69 λ clones hybridizable with one or more of the probes A, B and Q were obtained. From each of these clones, λDNA was prepared by the method as described above, treated with a restriction enzyme EcoRI and electrophoresed to thereby confirm the size of the DNA insert.

Example 5 Estimation of FcγBP mRNA Size

By using the probe Q obtained in Example 2, the molecular size of the mRNA of FcγBP was estimated. For comparison, use was made of the mRNAs of known proteins, i.e., Dystrophin mRNA of 14.0 kbp [M. Koenig et al. (1988) Cell 53:219-228] and Ryanodine Receptor mRNA of 15.2 kbp [F. Zarzato et al., (1990), J. Biol. Chem., 265:2244-2256]. As the cDNA probes for these comparative mRNAs, synthetic probes each having the base sequence reported in the reference were prepared and employed in polymerase chain reaction (PCR) to thereby give probes DYS and RDR respectively.

The Dystrophin mRNA and Ryanodine Receptor mRNA originated in human skeletal muscle polyadenylated RNA (manufactured by Clontech).

2 μg of the polyadenylated RNA obtained from colonic mucosal epithelial cells, 1 μg of the human skeletal muscle polyadenylated RNA or a mixture thereof was electrophoresed in the same manner as the one described in Example 2 (4) and thus transferred onto a nylon membrane. Then this nylon membrane was subjected to hybridization with the use of the probe Q followed by the detection by autoradiography.

To perform the hybridization of the comparative mRNAs on the same membrane, this nylon membrane was incubated together with 20 ml of a solution containing 50 mM of Tris-HCl buffer (pH 7.5), 1.25 mM of EDTA, 3×SSC, 1×Denhard's solution, 1% of SDS and 50% of formamide at 70° C. for 1 hour. Subsequently, it was washed by shaking in a washing liquor containing 0.2×SSC and 0.1% of SDS at room temperature for 10 minutes twice. Then the nylon membrane was subjected to autoradiography and thus it was confirmed that the band had been disappeared (dehybridization).

Next, hybridization was performed by using the probe DYS in the above-mentioned manner followed by the detection by autoradiography. This nylon membrane was subjected to dehybridization by the same method as the one described above and it was confirmed that the band had been disappeared. Finally, hybridization with the probe RDR was carried out in the same manner followed by the detection of the band by autoradiography.

On the basis of the results thus obtained, the mobilities of the bands of the Dystrophin mRNA and Ryanodine Receptor mRNA were each plotted against molecular size to thereby give a standard curve. Thus the molecular size of the mRNA of FcγBP was estimated as about 17 kbp form its mobility (FIG. 1).

Example 6 Identification of Base Sequence of cDNA Encoding FcγBP (1)

To identify the base sequence of the region encoding the amino acid sequence of the protein having the IgG Fc portion-binding activity, 5 necessary clones were selected by the following methods from among the 69 λ clones obtained in the above Example 4. Then the base sequence was identified by using a DNA sequencer (Model 373A, manufactured by Applied Biosystems).

(1) Clone X1

Among the cDNA clones obtained by the screening via hybridization with the probes A, B and Q, one which was hybridizable not with the probes Q and B but with the probe A alone was obtained. Then a fragment of about 700 bp, which was excised with EcoRI and SmaI at the opposite end to the probe A in the cDNA insert of this clone, was recovered and named probe X. Next, the cDNA library was screened by the same method as the one of Example 3 with the use of this probe X. Thus a clone X1 hybridizable with the probes X, A and B was obtained. The cDNA insert of this clone X1 was cleaved with EcoRI and electrophoresed on an agarose gel to thereby separate and recover a DNA of about 3,300 bp. Then this DNA was inserted into the EcoRI site of a plasmid vector pBluescript SK(+). Subsequently, the restriction map was formed and the base sequence was identified.

(2) Clone Y1

A fragment of about 800 bp, which was excised with EcoRI and SacI on the opposite side to the region containing the probe B in the cDNA of the clone X1, was recovered and named probe Y. Next, the cDNA library was screened by the same method as the one of Example 3 with the use of this probe Y. From among the clones thus obtained, clone Y1 was obtained as the one having the longest cDNA. The cDNA insert of this clone Y1 was cleaved with EcoRI and electrophoresed on an agarose gel to thereby separate and recover a DNA of about 1,900 bp. Then this DNA was inserted into the EcoRI site of a plasmid vector pBluescript SK(+). Subsequently, the restriction map was formed and the base sequence was identified.

(3) Clone C72

A fragment of about 150 bp, which was excised with SacI and SphI on the opposite side to the region containing the probe X in the cDNA of the clone Y1, was recovered and named probe Y150. Next, the cDNA library (cDNA size ranging from 2 to 4 kbp) was screened by the same method as the one of Example 3 with the use of this probe Y150 to thereby give 9 clones. From among the cDNA inserts of the clones thus obtained, a cDNA showing the longest extension on the opposite side to the Y region from Y150 and containing Y150 was obtained and named clone C72. The cDNA insert of this clone C72 was cleaved with EcoRI and electrophoresed on an agarose gel to thereby separate and recover a DNA of about 1,200 bp. Then this DNA was inserted into the EcoRI site of a plasmid vector pBluescript SK(+). Subsequently, the restriction map was formed and the base sequence was identified.

(4) Clone NZ4

A fragment of about 450 bp, which was excised with EcoRI and SacI on the opposite side to the region containing the probe Y150 in the cDNA of the clone C72, was recovered and named probe Z. By using an incubated cell line HT-29-18-N2 originating in human colonic cancer, a λgt10 cDNA library was constructed by the same method as the one described in Example 3 (2) to (5). Then this library was screened by the same method as the one of Example 3 with the use of the above-mentioned probe Z to thereby give 4 clones. From among the clones thus obtained, one containing the longest part not overlapping C72 was selected and named clone NZ4. The cDNA insert of this clone NZ4 was cleaved with NotI and electrophoresed on an agarose gel to thereby separate and recover a DNA of about 900 bp. Then this DNA was inserted into the EcoRI site of a plasmid vector pBluescript SK(+). Subsequently, the restriction map was formed and the base sequence was identified.

(5) Clone V11

The base sequences of the clones, which were hybridizable with all of the probes A, B and Q in the screening with the use of these probes, were analyzed to thereby give a clone having the same base sequence as the one of the A-B region located on the terminal side of the clone X1 which had been previously sequenced. This clone was named clone V11. The cDNA insert of this clone V11 was cleaved with EcoRI and electrophoresed on an agarose gel to thereby separate and recover a DNA of about 3,700 bp. Then this DNA was inserted into the EcoRI site of a plasmid vector pBluescript SK(+). Subsequently, the restriction map was formed and the base sequence was identified.

Example 7 Assumption of the Presence of Clone NZ4 in the Neighborhood of 5′ End of FcγBP mRNA

The 5 clones obtained in Example 6 were extended toward the 5′ end or the 3′ end in the order of NZ4-C72-Y1-X1-V11. When the base sequences of these clones were translated into amino acids, it was assumed that they would be extended toward the 5′ end. Thus a library constructed by random priming was screened with the use of the clone NZ4 DNA located to the uppermost stream at the present stage. As a result, 13 independent clones were obtained but none was extended to the 5′-upstream from NZ4. When the base sequence of NZ4 was translated into amino acids, the ATG codon corresponding to methionine seemingly located to the 5′-uppermost stream in the open reading frame was similar to Kozak's rule, which suggested that this ATG might be the initiation methionine. However, the first A in this ATG codon appeared early (i.e., located at the 9-position) in the clone NZ4 and no stop codon in frame existed in the preceding 9 bases. Therefore, it could not be denied that the cDNA sequence might be extended toward the 5′ (N) end not only in the transcription product but also at the translation level. Thus the following experiment was carried out in order to examine the transcription initiation site and to assume the translation initiation site by the primer extension method.

(1) Preparation of all RNAs and Polyadenylated RNA

Two primers, i.e., primer 1 (GCTGATAGTTCTGCAGGAAGGCTGTGAGGAATTCCTCTCTGCCAGTGTT-50 mer, SEQ ID NO:10) and primer 2 (GCTCCAGCCCAGAGTATCCACCAGCTCCATAGG-33 mer, SEQ ID NO:11) were synthesized with a DNA synthesizer (Model 394, manufactured by Applied Biosystems) and purified with an OPC column (manufactured by Applied Biosystems). 100 pmol of each primer was labeled with γ[³²P]ATP by using T4 polynucleotide kinase and purified with a Microspin™ S-200HR column (manufactured by Pharmacia). 0.5 pmol of each primer was used in each reaction.

(3) Primer Annealing and Extension Reaction

All RNAs (20 μg) and polyadenylated RNA (2.5 μg) originating in human colonic mucosal epithelial cells and HT-29-18-N2 were each mixed with the primers in an annealing buffer [10 mM of Tris-HCl (pH 7.5), 1 mM of EDTA, 250 mM of KCl] and thermally denatured by heating at 95° C. for 5 minutes. Then hybridization was carried out by incubating at 58° C. for 1 hour and then at room temperature or 37° C. for 1.5 hours. Subsequently, the extension reaction was performed in the following manner. An annealing sample was precipitated from ethanol and the obtained precipitate was dissolved in RTase buffer [33 mM of Tris-HCl (pH 8.3), 20 mM of KCl, 13.3 mM of MgCl₂, 13.3 mM of DTT, 0.33 mM of dNTPs, 50 μg/ml of actinomycin D]. After adding 20 U of a reverse transcriptase (RnaseH-free MMLV RTase, manufactured by Toyobo), the mixture was incubated at 42° C. for 1 hour. After the completion of the reaction, the mixture was treated at 95° C. for 3 minutes to thereby inactivate the RTase. Then RNase A was added to give a concentration of 10 μg/ml and the mixture was incubated at 37° C. for 30 minutes to thereby decompose the template RNA. After extracting successively with phenol/chloroform and chloroform and precipitating from ethanol, the precipitate was electrophoresed on a 5% sequence gel. After the completion of the electrophoresis, the gel was treated with a fixing solution (10% of acetic acid, 15% of methanol), dried and then subjected to autoradiography. As a marker, use was made of M13mp18 which had been reacted with Sequenase ver 2.0 DNA Sequencing Kit (manufactured by Toyobo).

Thus the following results were obtained. As the result of the extension with the primer 1 by using each all RNA specimen as a template, a strong band was observed at around the base of the 118-position from the primer. As the result of the extension with the primer 1 by using each polyadenylated RNA specimen as a template, a weak extension band was observed at around the base of the 157-position, in addition to the one at around the 118-position. The 5′ end of the NZ4, which was considered to be located at the 5′-uppermost stream, was referred to as +1 for convenience. Then these bands corresponded respectively to +27 and −13. The extension stopped at this +27 position. This is seemingly because a palindromic structure suggesting the formation of the secondary structure at round the 5′ end of NZ4 might occur. In the case of the primer 2 which was located 5′-upstream and had been constructed to minimize the formation of such secondary structure, in fact, a broad band was observed from −10 to −16 and a weak single band was further detected at the position corresponding to −23. No band was detected upstream from −23.

These results indicate that the transcription initiation site is located upstream from the 5′ end of NZ4 by 10 to 20 bases. When no ATG codon in frame is involved in this range, there is a strong possibility that the ATG located at the 5′-uppermost stream at the present stage in ORF would be the translation initiation site. These facts point out that the clone NZ4 is extremely close to the N end in the order of NZ4-C72-Y1-X1-V11.

Example 8 Construction of Expression cDNA/vector System (A: Preparation of partial cDNA employed in expression)

For the expression of the protein, the λDNA clones (#NZ4, #C72, #Y1, #X1 and #V11) having partial cDNA of FcγBP inserted therein were cleaved with EcoRI or NotI and the cDNA inserts were subcloned into a cyclic plasmid pBluescript SK(+). The plasmids thus obtained were respectively named pNZ4, pC72, pY1, pX1 and pV11.

(1) pNZ4

The λ clone (#NZ4) was completely digested with NotI. Then the insert of about 900 bp was separated and purified by agarose gel electrophoresis and then ligated to the NotI site of pBluescript SK(+). Then selection was made of a clone in which the 5′→3′ direction of the protein code strand of the cDNA had been inserted in the opposite direction to the lacZ gene of the plasmid. The entire base sequence of the insert is shown in SEQ ID NO:1.

In Sequence Listing in the present invention, base sequences originating in cDNAs are given in capital letters, those originating in pBluescript SK(+) are given in small letters, and those originating in synthetic adaptors and synthetic oligonucleotides are given in underlined small letters.

Each amino acid sequence given in Sequence Listing is one which is assumed on the basis of the base sequence by the universal codon while referring ATG which agrees with Kozak's sequence as to the initiation codon.

(2) pC72

The λ clone (#C72) was completely digested with EcoRI. Then the insert of about 1,300 bp was separated and purified by agarose gel electrophoresis and then ligated to the EcoRI site of pBluescript SK(+). Then selection was made of a clone in which the 5′→3′ direction of the cDNA had been inserted in the opposite direction to the lacZ gene of the plasmid. The entire base sequence of the insert is shown in SEQ ID NO:2.

(3) pY1

The λ clone (#Y1) was completely digested with EcoRI. Then the insert of about 1,900 bp was separated and purified by agarose gel electrophoresis and then ligated to the EcoRI site of pBluescript SK(+). Then selection was made of a clone of the same size. The entire base sequence of the insert is shown in SEQ ID NO:3.

(4) pX1

The λ clone (#X1) was completely digested with EcoRI. Then the insert of about 3,300 bp was separated and purified by agarose gel electrophoresis and then ligated to the EcoRI site of pBluescript SK(+). Then selection was made of a clone of the same size. The entire base sequence of the insert is shown in SEQ ID NO:4.

(5) pV11

The λ clone (#V11) was completely digested with EcoRI. Then the insert of about 3,700 bp was separated and purified by agarose gel electrophoresis and then ligated to the EcoRI site of pBluescript SK(+). Then selection was made of a clone of the same size. The entire base sequence of the insert is shown in SEQ ID NO:5.

Example 9 Construction of Expression cDNA/vector System (B: Ligation of partial cDNA for expression of protein)

(1) Preparation of pNZC7

The plasmid pNZ4 (5 μg) having the cDNA clone inserted therein was completely digested with restriction enzymes XhoI and BglII (each 50 U) and electrophoresed on a low melting agarose gel. Then a fragment of about 400 bp containing the 5′ end of the cDNA of FcγBP was separated, extracted with phenol and recovered by precipitation from ethanol (fragment 1). Next, the second plasmid pC72 (5 μg) was completely digested with XhoI and BglII. Then a fragment of about 4.2 kbp containing the vector part was isolated by electrophoresis in the same manner as the one described above (fragment 2). The fragments 1 and 2 were each dissolved in 10 μl of TE buffer. 2 μl portions of these solutions were mixed with 16 μl of the solution A (DNA Ligation Kit, manufactured by Takara Shuzo) and 4 μl of the solution B and incubated at 16° C. for 30 minutes to thereby perform ligation. By 5 μl of this mixture was transformed 100 μl of competent E. coli (XL1-Blue) which was then incubated on an LB plate containing 100 μg/ml of ampicillin at 37° C. overnight. From the colonies thus formed, the plasmid DNA was purified to thereby give a plasmid pNZC7 wherein the fragment 1 had been ligated to the fragment 2.

(2) Preparation of fragment 5

pNZC7 (5 μg) was completely digested with 50 U portions of XhoI and BstXI and a fragment of about 1,300 bp was recovered by electrophoresis (fragment 3). The third plasmid pY1 (5 μg) was completely digested with 50 U portions of BstXI and HincII and a fragment of about 420 bp was recovered by electrophoresis (fragment 4). Then these fragments 3 and 4 were ligated to each other with DNA ligase by the same method as the one described above and electrophoresed. Thus a fragment of about 1,750 bp, in which the above fragments (each 1 mole) had bound to each other at the BstXI site, was recovered (fragment 5).

(3) Preparation of pXV2

The fourth plasmid pX1 (5 μg) was completely digested with 50 U portions of HincII and BamHI and a fragment of about 2,780 bp was recovered by electrophoresis (fragment 6). The fifth plasmid pV11 (5 μg) was completely digested with 50 U of BamHI and a fragment of about 3,350 bp was recovered by electrophoresis (fragment 7). Then these fragments 6 and 7 were ligated to each other with DNA ligase by the same method as the one described above and electrophoresed. Thus a fragment of about 6,100 bp, in which the above fragments (each 1 mole) had bound to each other, was recovered (fragment 8). This fragment 8 was then ligated to pBluescript SK(+), which had been digested with HincII and BamHI, with DNA ligase and then competent E. coli was transformed thereby. From the transformants thus obtained, plasmids were recovered and the base sequence of each plasmid was identified. Thus a plasmid clone containing the fragment 8, in which the fragments 6 and 7 had been ligated in the correct direction, was obtained. The one in which the 5′→3′ direction of the fragment 8 had been inserted in the opposite direction to the lacZ gene of the plasmid was referred to as pXV2.

(4) Preparation of pNV11

pXV2 was completely digested with XhoI and HincII and a fragment (about 9.1 kbp) containing the vector was recovered by electrophoresis (fragment 9). This fragment 9 was ligated to the above-mentioned fragment 5 with the use of DNA ligase and then competent E. coli (XL1-Blue) was transformed thereby. From the transformants thus obtained, plasmid pNV11 (about 10.8 kbp) containing a cDNA of about 7.8 kbp (fragment 10) was obtained.

(5) Synthesis of Oligonucleotide Adaptor Containing Stop Codon (Corresponding to UAG)

By using a DNA synthesizer (model 394, manufactured by Applied Biosystems), the following oligonucleotides, which had three TAG differing in frame and NotI and SpeI sites at both ends, were synthesized: (1) 5′-CTA GTT AGT TAG TTA GGG TAC CGC-3′, SEQ ID NO:12; and (2) 5′-GGC CGC GGT ACC CTA ACT AAC TAA-3′ SEQ ID NO:13. 10 nmol portions of the oligonucleotides 1 and 2 were mixed together (146 μl in total), heated at 95° C. for 1 minutes and at 85° C. for 10 minutes and then gradually cooled to 40° C. at a rate of 0.33° C./min to thereby prepare an adaptor containing the stop codon (TA-III adaptor). Then the 5′ end of this adaptor (2.1 nmol) was phosphorylated by the standard method with the use of ATP and polynucleotide kinase.

0.83 pmol of the pBluescript SK(+) vector was completely digested with NotI and SpeI and then mixed with 250 pmol of the phosphorylated TA-III adaptor and ligated thereto by incubating at 16° C. for 30 minutes with the use of a DNA ligation kit. After precipitating from ethanol, the precipitate was completely digested with NotI in 50 μl of reaction volume in such a manner as to give the adaptor sequence once. Next, a band of about 3 kbp was recovered by low melting agarose gel electrophoresis, extracted with phenol and precipitated from ethanol. The precipitate thus obtained was subjected to autoligation with the use of a DNA ligation kit and then competent E. coli (XL1-Blue) was transformed thereby. Among the plasmids obtained from the colonies formed by incubating the transformant on an LB plate containing ampicillin overnight, selection was made of a plasmid into which the TA-III adaptor had been inserted (pBLS/TAIII).

(6) Preparation of pNV11-ST

5 μg of the plasmid pNV11 was completely digested with SpeI and a fragment of about 7.8 kbp was recovered by electrophoresis and dissolved in 10 μl of TE buffer (fragment 11). 2 μg of the plasmid (pBLS/TAIII) was completely digested with SpeI and its end was dephosphorylated with bacteria alkaline phosphatase (2 U). Then it was treated with phenol/chloroform twice and precipitated from ethanol. The precipitate was dissolved in 10 μl of TE buffer (fragment 12). Next, 2 μl portions of the fragments 10 and 11 were mixed together and ligated with the use of a DNA ligation kit (manufactured by Takara Shuzo). Then competent E. coli (XL1-Blue) was transformed thereby and incubated on an LB plate containing ampicillin overnight. Among the colonies thus formed, one in which the TA-III adaptor had been ligated on the 3′ side of the inserted cDNA was selected by analyzing the restriction map and the base sequence. Thus the clone pNV11-ST was obtained.

The E. coli strain containing the above-mentioned plasmid pNV-ST has been deposited as Escherichia coli XL1-Blue [pNV11-ST] at National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology (1-3 Higashi 1-chome, Tsukuba-Shi Ibaraki-Ken 305, Japan) under the accession number FERM BP-4625 in accordance with Budapest Treaty since Apr. 1, 1994.

FIG. 2 shows the relations among the partial cDNA (clone pNV11-ST; about 7.8 kbp) employed in the expression of FcγBP and the clones NZ4, C72, Y1, X1 and V11 described in Example 6 and employed in the construction thereof.

Example 10 Construction of Expression cDNA/Vector System (C: Integration into protein expression vector)

(1) Preparation of pcDL-SRα/NOT Vector

To enable the cDNA integration, the restriction enzyme sites of a vector pcDL-SRα296 (kindly afforded by Dr. Yutaka Takebe, National Institute of Health; hereinafter sometimes referred to simply as SRα) were altered. First, 2 μg of SRα was completely digested with EcoRI and precipitated from ethanol. The precipitate was dissolved in Klenow buffer [70 mM of Tris-HCl (pH 7.5), 1 mM of EDTA, 200 mM of NaCl, 70 mM of MgCl₂, 1 mM portions of dATP, dCTP, dGTP and dTTP] and incubated with 0.4 U of Klenow fragment at 37° C. for 15 minutes to thereby blunt-ended the plasmid. After the precipitation from ethanol, the precipitate was dissolved in TE buffer and an NotI linker, which had been phosphorylated at the 5′ end, was ligated thereto with the use of a DNA ligation kit. After the precipitation from ethanol, the precipitate was completely digested with NotI and electrophoresed on an agarose gel. Thus a DNA of about 3.7 kbp was excised and recovered by extracting with phenol. The DNA thus recovered was subjected to autoligation by using a DNA ligation kit and then competent E. coli (XL1-Blue) was transformed thereby. Next, the target plasmid (pcDL-SRα/NOT) digested not with EcoRI but with NotI was selected.

(2) Insertion of Expression cDNA

The protein expression vector (pcDL-SRα/NOT) was completely digested with NotI and KpnI and a fragment of about 3.7 kbp was recovered by electrophoresis (fragment A).

The cDNA insertion vector (pNV11-ST) was completely digested with NotI and KpnI and a fragment of about 7.8 kbp was recovered by electrophoresis (fragment 13). The entire base sequence of this fragment 13 is shown in SEQ ID NO:6.

This base sequence and the amino acid sequence deduced therefrom were retrieved with GenBank Rel. 80. Thus it was confirmed that the base sequence and the amino acid sequence are both novel ones.

The fragment A and the fragment 13 were ligated by using a DNA ligation kit and then competent E. coli (XL1-Blue) was transformed thereby. From among the colonies formed by incubating the transformant on an LB plate containing 100 μg/ml of ampicillin, a plasmid having the fragment 13 inserted therein was selected by cleaving with restriction enzymes. Thus the clone pNV11-SR was obtained.

Example 11 Expression of Partial cDNA of FcγBP in COS7 Cell

(1) Recovery of Expression cDNA/Vector from Escherichia coli

The Escherichia coli transformed by the FcγBP cDNA expression plasmid (pNV11-SR) obtained in Example 10 was incubated in 10 ml of LB medium at 37° C. under shaking overnight. Next, the culture medium was added to 500 ml of LB medium and shaking was continued until OD₆₀₀ reached 0.8. When OD₆₀₀ of 0.8 was attained, 2.5 ml of a chloramphenicol solution (34 mg/ml) was added thereto and the mixture was incubated overnight. After separating the cells by centrifugation, the plasmid DNA was prepared by the alkali method in the conventional manner. The plasmid was purified by ultracentrifuging (90,000 rpm, 3 hours) under density gradient of cesium chloride twice and dialyzing against TE buffer and then employed in the expression of the protein.

(2) Transfection into COS7 Cell

Transfection was performed in the following manner in order to examine the properties of the protein through the tentative expression of the plasmid vector (pNV11-SR), which had the partial cDNA (abut 7.8 kbp) of FcγBP integrated therein, in COS7 cells. 2×10⁷ COS7 cells were added to a dish of 35 mm in diameter and incubated in RPMI 1640 medium (0.2% of sodium hydrogencarbonate, 10 U/ml of penicillin, 0.01% of streptomycin) containing 10% of FBS overnight. On reaching 40-60% confluence, the cells were washed with serum-free RPMI 1640 medium twice.

10 μg of the plasmid pNV11-SR dissolved in 250 μl of RPMI 1640 medium was mixed with 10 μl of a lipofection reagent (Transfectam, manufactured by Sepracor) dissolved in 250 μl of RPMI 1640 medium and the obtained mixture was immediately added onto the COS7 cells. After incubating at 37° C. for 6 hours, the medium was removed followed by the addition of 2 ml of RPMI 1640 medium containing 10% of serum. Then incubation was carried out at 37° C. under 5% of CO₂ for 2 days.

(3) Confirmation of Expressed Protein

The dish (diameter: 35 mm) wherein the COS7 cells transfected with pNV11-SR had been incubated was washed with 2 ml of PBS (−) twice. Then 2 ml of 99.5% ethanol was added and the cells were fixed at room temperature for 5 minutes and then washed with 2 ml of PBS (−) twice. One ml portion of culture supernatants of hybridomas producing the monoclonal antibodies (K9 and K17) against FcγBP employed in screening the λ phage were added. Each mixture thus obtained was incubated at room temperature for 1 hour and washed with PBS (−) thrice. Then horse radish peroxidase (HRP)-labeled goat antimouse IgG (H+L) F(ab′)₂ fragment (manufactured by Zymed) was added thereto followed by incubation at room temperature for 30 minutes. After washing 2 ml of PBS (−) thrice, a 0.036% aqueous solution of hydrogen peroxide and a 0.1% solution of diaminobenzidine in 0.1 M Tris HCl (pH 7.2) were added at a ratio of 1:1 so as to effect color development (room temperature, 10 minutes). Thus the cells wherein the protein was expressed were confirmed. As a control, use was made of a sample to which no primary antibody but (HRP)-labeled goat antimouse IgG (H+L) F(ab′)₂ fragment alone was added as the secondary antibody.

FIG. 3 shows the results. When the culture supernatant of the hybridomas producing the K9 monoclonal antibody (FIG. 3, A) or that of the hybridomas producing the K17 monoclonal antibody (FIG. 3, B) was added, cells specifically reacting therewith were observed in each case. On the other hand, no reactive cell was observed in the control (FIG. 3, C).

Example 12 Detection of Ability of Recombinant Protein to Bind to Human IgG and its Properties

(1) Confirmation of Binding of Human IgG

The COS7 cells (in a dish of 35 mm in diameter) transfected with the plasmid pNV11-SR, into which the partial cDNA of FcγBP had been integrated, were washed with PBS (−) twice. Then 2 ml of 99.5% ethanol was added thereto and the cells were fixed at room temperature for 5 minutes and then washed with 2 ml of pBS (−) twice. Next, a human IgG fraction (manufactured by Cappel) purified by affinity chromatography was diluted with RPMI 1640 medium containing 10% of FBS so as to give a concentration of 10 μg/ml. One ml of the obtained dilution was added to the dish and incubated at room temperature for 1 hour. After, washing with 2 ml of PBS (−) thrice, it was incubated with HRP-labeled goat antihuman IgG F(ab′)₂ fraction (#109-D36-088, manufactured by Cosmo Bio) at room temperature for 30 minutes. After washing 2 ml of PBS (−) thrice, a mixture (1:1) of a 0.036% aqueous solution of hydrogen peroxide and a 0.1% solution of diaminobenzidine in 0.1 M Tris HCl (pH 7.2) was added so as to effect color development (room temperature, 10 minutes). Thus the binding of IgG to the expressed protein was confirmed.

(2) Specific Binding of IgG

The COS7 cells (in a dish of 35 mm in diameter) transfected with pNV11-SR were washed with 2 ml of PBS (−) twice. Then 2 ml of 99.5% ethanol was added thereto and the cells were fixed at room temperature for 5 minutes and then washed with 2 ml of pBS (−) twice.

Next, HRP-labeled human IgG fraction (#55902, manufactured by Cappel) purified by affinity chromatography was diluted with RPMI 1640 medium containing 10% of FBS so as to give a concentration of 10 μg/ml (solution 1). To this solution 1, were added the following immunoglobulins (50 μg/ml) as a competitive inhibitor.

(1) Human IgG fraction purified by chromatography (#55908, manufactured by Cappel).

(2) Human IgG Fc fraction purified by chromatography (#55911, manufactured by Cappel).

(3) Human IgG F(ab′)₂ fraction purified by chromatography (#55910, manufactured by Cappel).

(4) Human IgM fraction purified by chromatography (#55916, manufactured by Cappel).

(5) Human serum IgA purified by chromatography (#55906, manufactured by Cappel).

(6) Human secretor IgA purified by chromatography (#55905, manufactured by Cappel).

The above competitive inhibitors (1) to (6) were separately added to the solution 1. One ml of each solution thus obtained was added to the dish in which the cells had been fixed followed by incubation at room temperature for 1 hour. After washing with 2 ml of PBS (−) thrice, a mixture (1:1) of a 0.036% aqueous solution of hydrogen peroxide and a 0.1% solution of diaminobenzidine in 0.1 M Tris HCl (pH 7.2) was added so as to effect color development (room temperature, 10 minutes). Thus the binding of IgG to the expressed protein was examined. As a control, use was made of the solution 1 containing no competitive inhibitor.

FIGS. 4 and 5 show the results. When no competitive inhibitor was added (i.e., the control), the cells were stained (FIG. 4, A). When the purified human IgG fraction (FIG. 4, B) and the purified human IgG Fc fraction (FIG. 4, C) were added, however, the cells were not stained. On the other hand, addition of the IgG F(ab′)₂ fraction (FIG. 5, D), the human IGM fraction (FIG. 5, E), the human serum IgA (FIG. 5, F) and the human secretor IgA (FIG. 5, G) could not inhibit the binding of the HRP-labeled human IgG. These facts indicate that FcγBP binds specifically to IgG Fc in human antibodies.

Example 13 Tissue-specificity of the Expression of FcγBP mRNA

To examine the specificity of the expression of FcγBP in human tissues, the expression of the mRNA was analyzed by Northern blotting. A nylon membrane, on which 2 μg portions of polyadenylated RNAs purified from human heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas had been blotted (Human Multiple Northern Blots, #7760-1, manufactured by Clontech), was subjected to prehybridization under the same conditions as those employed in Example 2 (4) followed by hybridization with the use of the probe Y labeled with [³²P]. After washing, bands were detected by autoradiography. After performing the autoradiogram at −80° C. for 2 days, a band of about 17 kbp was detected in the placenta (FIG. 6), while other tissues showed negative results. It was therefore assumed that the FcγBP protein had been expressed in the placenta.

Example 14 Northern Blotting Analysis with 3 Different Probes

The mRNA extracted from colonic mucosal epithelial cells was subjected to Northern blotting by using the probes Q and A obtained in Example 2 (3) and the probe Y obtained in Example 6 (2) in order to confirm that these probes would be hybridizable with the same mRNA.

15 μg of all RNAs extracted from colonic mucosal epithelial cells by the AGPC method were dissolved in 4.5 μl of sterilized water, mixed with 2 μl of 5×MOPS buffer, 3.5 μl of formaldehyde and 10 μl of formamide and then thermally denatured at 60° C. for 15 minutes followed by electrophoresis on a 1% agarose gel in the presence of formaldehyde. After the completion of the electrophoresis, the RNAs were transferred overnight onto a nylon membrane (Biodyne A, manufactured by Pall Corp.) by the capillary method. After fixing the RNAs to the nylon membrane by UV crosslinking, prehybridization was performed in 10 ml of a hybridization solution (5×SSPE, 5×Denhardt's solution, 50% of formamide, 0.5% of SDS, 100 μg/ml of thermally denatured salmon sperm DNA) at 42° C. for 8 hours.

Subsequently, the probes A, Q and Y were each radioisotope-labeled with α[³²P]dCTP by using Megaprime Labeling Kit (manufactured by Amersham). 1×10⁸ dpm of each probe was added together with 5 ml of the hybridization solution to the nylon membrane which had been subjected to the prehybridization. After sealing, hybridization was carried out at 42° C. overnight. Then the nylon membrane was washed in a solution containing 0.2×SSC and 0.2% of SDS at 65° C. for 40 minutes thrice. Then the nylon membrane was dried and exposed to an X-ray film overnight.

As a result, a band of approximately 17 kbp was detected by using each of the probes A, Q and Y. which proves that these 3 probes are hybridizable with the same mRNA from the viewpoint of molecular weight (FIG. 7).

Example 15 Identification of Base Sequence of cDNA Encoding FcγBP (2)

Examples 4 and 6 state the identification of the partial base sequence [i.e., about 7.8 kbp (7,826 bases) starting from the 5′ end] of the cDNA encoding the amino acid sequence of the protein capable of binding to the Fc region of IgG. Now, the method for identifying the residual base sequence (about 8.6 kbp) will be illustrated.

(1) Structure and Classification of cDNA

The cDNA clones obtained by the screening via the hybridization with the use of the probe A, B or Q in Example 4 were each amplified in Escherichia coli and then mapped with the use of the probes A, B and Q. As a result, it was found out that each of these clones was hybridizable with at least one of the 3 probes and the sites homologous with the probes were located in the order of A→B, B→Q or Q→A on the cDNAs.

Based on these results, it was assumed that the FcγBP gene has a structure wherein a unit consisting of the sequences homologous with the probes A, B and Q linked together in the order of A→B→Q is repeated in tandem. In cDNAs clone hybridizable with the probe B, a part (about 280 bp) of the base sequence of the probe B was amplified by PCR with the use of the following primers.

Primer (P-1):

5′-GCC TGC GTG CCC ATC CAG-3′ SEQ ID NO:14.

Primer (P-2):

5′-CTC ATA GTT GGG CAG CGAC-3′ SEQ ID NO:15.

The fragments amplified by PCR were separated by agarose gel electrophoresis and collected from the gel followed by the analysis of the base sequences. Based on these base sequences, the cDNA clones hybridizable with the probe B were classified into the following 3 groups, while the cDNA clones not hybridizable with the probe B were referred to as the group 4.

Group 1: cDNA clones having the same sequence as that of the amplified fragment of the clone V11.

Group 2: those suffering from the replacement of 5 bases, compared with the sequence of the group 1, and containing no HincII site in the fragment.

Group 3: those suffering from the replacement of 7 bases, compared with the sequence of the group 1, and containing the HincII site in the fragment.

Group 4: those not hybridizable with the probe B.

(2) Clone T5

To isolate the cDNA having poly A moiety on the 3′ side, the cDNA library constructed in Example 3 with the use of the oligo dT primer was screened by using the probe A, B or Q in the same manner as the one described in Example 4. As a result, hybridization occurred exclusively with the probe Q. Among the cDNA clones thus obtained, the one having the longest cDNA insert was referred to as clone T5.

The cDNA insert of the clone T5 was cleaved with EcoRI and separated and purified by agarose gel electrophoresis followed by the insertion into the EcoRI site of the plasmid vector pBluescript SK(+). Subsequently, the restriction map of T5 was formed and the entire base sequence thereof was identified.

In the cDNA insert of the clone T5, a region of about 550 bp between the BamHI site about 1 bp apart from poly (A⁺) on the 5′ side and the PstI site about 1.6 kbp apart from the poly (A⁺) on the 5′ side was referred to as probe V. This probe V was not hybridizable with the clones NZ4, C72, Y1, X1, V11, A53, A40 and A 31 but specific to T5.

(3) Clone A43

The cDNA clones hybridizable with the probe A, B or Q were subjected to hybridization by using the probe V in the same manner as the one described in Example 4 (2) to thereby give a clone which was hybridizable not with the probes A and B but with the probes V and Q. The cDNA insert of this clone was cleaved with EcoRI and separated and purified by agarose gel electrophoresis followed by the insertion into the EcoRI site of the plasmid vector pBluescript SK(+). Subsequently, the restriction map was formed and the entire base sequence thereof was identified.

As the result of the analysis of the base sequence, it was confirmed that the sequence of about 2 kbp containing the region hybridizable with the probe Q agreed with the sequence of the part overlapping T5.

(4) Clone A8

To obtain a cDNA extended toward the 5′ end from the clone A43, synthesis was made of the following primers by which the base sequence (about 240 bp) around the 5′ end of the clone A43 could be amplified.

Primer (P-3):

5′-TGT TGG GAC GAA TGT CGG-3′ SEQ ID NO:16.

Primer (P-4):

5′-TCA CAG CCA ACC TGT GCC-3′ SEQ ID NO:17.

The cDNA clones of the groups 1, 2 and 3 as classified in Example 15 (1) were subjected to PCR with the use of the above-mentioned primers (P-3) and (P-4). The fragments amplified by PCR were separated by agarose gel electrophoresis and collected followed by the analysis of the base sequences. Thus a cDNA clone having a PCR fragment of the same base sequence as that of A43 was selected from among the cDNA clones of the group 3 as classified in Example 15 (1). This clone was referred to as clone A8. The entire base sequence of the clone A8 was analyzed and it was thus confirmed that the base sequence on the 3′ side was completely identical with the overlapping sequence on the 5′ side of A43.

(5) Clones A53 and A40

From among the clones falling within the group 2, 2 different clones each having a region hybridizable with the probes Q and A on the 5′ side were selected on the basis of the restriction maps. Between these clones thus selected, one which had the longer region overlapping the 3′ side of the clone V11 was referred to as clone A53, while another one having the shorter region was referred to as clone A40.

The entire base sequences of the clones A53 and A40 were analyzed. Thus it was confirmed that a region on the 3′ side of the clone A53 (about 2.4 kbp) and a region on the 5′ side of the clone A40, which overlapped each other, had the same base sequence. Further, a comparison between a region (about 1.8 kbp) on the 3′ side of V11 and a region on the 5′ side of A53 overlapping each other indicated that the base sequences of these regions were completely consistent with each other except one base (i.e., the base at the position 6273: A in V11, G in A53).

(6) Clone A31

To screen a cDNA extended toward the 3′ end from the clone A40, the following procedure was performed. In the clones A53 and A40, the base sequences of the fragments amplified by the primers (P-3) and (P-4) were identical with neither the sequence of V11 of the group 1 nor the sequence of A8 of the group 3. Thus screening was carried out in the following manner with the use of these sequences as indicators. Namely, cDNA clones hybridizable with the probe A, B or Q, from among the cDNA clones (69 in total) other than those belonging to the groups 1 and 3, were subjected to PCR by using the primers (P-3) and (P-4) and the base sequences of the fragments thus amplified were identified. Then cDNA clones having the same base sequence as those of the clones A53 and A40 were selected. From among these clones, one having the PCR-amplified sequence on the 5′ side and extended toward the 3′ end was selected and referred to as clone A31.

By analyzing the entire base sequence of the clone A31, it was confirmed that the sequence on the 5′ side of A31 was identical with the overlapping region on the 3′ side of the clone A40, while the sequence on the 3′ side of A31 was identical with the overlapping region on the 5′ side of the clone A8.

Example 16 Identification of Base Sequence of cDNA Encoding FcγBP (3)

The cDNA of FcγBP had a structure of 16.4 kbp in full length in which a sequence consisting of a unit of 3.5 kbp (containing regions homologous with the probes A, B and Q) was repeated thrice in tandem and the repeating sequences had a homology of at least 95% with each other (FIG. 8).

As described above, the cDNAs employed in the identification of the base sequences in this repeating structure had been cloned depending on the strong reactivities with the probes A, B and Q. By comparing these base sequences to each other, it was confirmed that the base sequences of overlapping regions were identical and thus the relationships among the cDNA were clarified. Consequently, it was proved that a series of DNA fragments originated in a single mRNA (gene). To confirm this fact again, a cDNA fragment in a repeating structure different from pNV11SR containing the 5′ terminal cDNA, which had been already employed in expression, was made to undergo protein expression. Then it was examined whether or not the protein thus expressed would be recognized by the monoclonal antibodies K9 and K17 capable of recognizing FcγBP.

(1) Synthesis of Adaptor Containing Initiation Codon

To express each cDNA to be inserted into a vector from the 5′ end in full length, it is required to ligate the initiation codon (ATG) to the 5′ end of the cDNA. In order to achieve the translation of the protein within the same frame as that of FcγBP, it is furthermore required to regulate the frame between the initiation codon (ATG) and the 5′ end of the cDNA. Thus oligonucleotides for an adaptor satisfying the following conditions were synthesized.

Each synthetic oligonucleotide contains a base sequence consisting of 7 bases (GCCATGG), which is the same as the sequence of the initiation region of pNV11SR containing the inherent initiation codon (ATG) of FcγBP, and is consistent with Kozak's rule.

To facilitate the insertion into the vector, the HindIII site and the EcoRI site were added respectively to the 5′ side and 3′ side of this oligonucleotide. In order to regulate the frame, the following 3 oligonucleotides (FR-1S, FR-2S and FR-3S) were constructed. Furthermore, oligonucleotides FR-1A, FR-2A and FR-3A complementary respectively to the above-mentioned oligonucleotides were synthesized.

Oligonucleotide for adaptor

FR-1S: 5′-A GCT TCT GCA GCC ATG GG-3′ SEQ ID NO:18

FR-1A: 3′-AGA CGT CGG TAC CCT TAA-5′ SEQ ID NO:19

FR-2S: 5′-A GCT TCT GCA GCC ATG GGG-3′ SEQ ID NO:20

FR-2A: 3′-AGA CGT CGG TAC CCC-5′ SEQ ID NO:21

FR-3S: 5′-A GCT TCT GCA GCC ATG GGA G-3′ SEQ ID NO:22

FR-3A: 3′-AGA CGT CGG TAC CCT CTT AA-5′ SEQ ID NO:23

Each oligonucleotide was synthesized by using a DNA synthesizer (model 1395, manufactured by ABI) and purified.

Next, FR-1S, FR-2S and FR-3S were annealed respectively with FR-1A, FR-2A and FR-3A in the same manner as the one described in Example 9 (5) followed by the phosphorylation of the nucleotide at the 5′ end. The adaptors thus formed were referred to respectively as adaptors FR-1, FR-2 and FR-3.

(2) Alteration of TAIII/SK Vector

The XbaI site was added to the vector pBLS/TAIII containing the stop codon, which was constructed in Example 9 (5), in the following manner.

Namely, pBLS/TAIII was completely digested with a restriction enzyme XhoI by the same method as the one of Example 10 (1) and then blunt-ended. After ligating an XbaI linker having a phosphorylated end thereto, it was digested with XbaI again and autoligated. Then competent E. coli was transformed thereby and thus a vector pBLS/TAIII2 having the XbaI site inserted into the XhoI site of pBLS/TAIII was obtained.

(3) Insertion of Oligonucleotide Containing Initiation Codon

The pBLS/TAIII2 was completely digested with EcoRI and HindIII and electrophoresed on an agarose gel to thereby recover a vector part of about 3 kbp. Then the adaptors FR-1, FR-2 and FR-3 were each inserted into the above-mentioned pBLS/TAIII2 in the same manner as the one of Example 9 (5). By analyzing the base sequences, plasmids each having one adaptor (FR-1, FR-2 or FR-3) correctly inserted thereinto were obtained and referred to respectively as Fr1-SK2, Frs-SK2 and Fr3-SK2.

(4) Insertion of cDNA Fragment of FcγBP

To express the cDNA clone A53 in the same frame as that of FcγBP, the cDNA part of A53 was excised with EcoRI and inserted into the EcoRI site of Fr3-SK2. Then a plasmid in which the 5′ end of cDNA was located on the initiation codon side of Fr3-SK2 was selected and referred to as piF-A53.

Similarly, the cDNA insert of the clone A8 was inserted into Fr2-SK2 to thereby give piF-A8.

The piF-A53 and piF-A8 thus obtained were purified in the same manner as the one described in Example 11 (1) and then employed in an experiment of protein expression.

(5) Expression of piF-A53 and piF-A8

piF-A53 and piF-A8 were transfected into COS7 cells in the same manner as the one of Example 11-(2). After incubating for 2 days, the cells were stained by using monoclonal antibodies (K9/K17 mixture) by the same method as that of Example 11 (3) to thereby detect the proteins expressed tentatively. As a result, the cells transfected with piF-A53 and those transfected with piF-A8 were both stained with the monoclonal antibodies, which indicated that these cDNAs encoded some part of the protein recognized by either or both of the monoclonal antibodies K9 and K17 (FIGS. 9A and B). Thus it has been proved that A53 and A8 are each a portion of the repeated sequence of the entire cDNA of FcγBP.

Example 17 Assumption of the Presence of Clone NZ4 in the Neighborhood of 5′ End of FcγBP mRNA (2)

Based on the results of the primer extension carried out in Example 7, it was assumed that the transcription initiation site might be located within about 20 bases upstream from the clone NZ4 which was located at the 5′-uppermost stream at the present stage; and that the ATG at the 9-position in NZ4 might be the translation initiation site. To examine whether or not ATG in frame or the stop codon was located upstream from NZ4, therefore, the FcγBP gene was isolated from the genomic DNA library and its partial base sequence was identified.

(1) Genomic DNA Library

As the library, use was made of a commercially available one originating in human leukocytes (Vector, EMBL3 SP6/T7, manufactured by Clontech).

(2) Probe

As the probes to be employed in the screening, use was made of one prepared by excising the cDNA clone NZ4 from the vector with BamHI and the synthetic oligonucleotide employed in Example 7 (primer 2: (GCTCCAGCCCAGAGTATCCACCAGCTCCATAGG-33 mer, SEQ ID NO:11) respectively labeled with α[³²P] dCTP and γ[³²P] ATP by random prime labeling (NZ4) or end labeling (primer 2).

(3) Screening

The screening with the probe NZ4 was performed in accordance with the method for screening a cDNA library as described in Example 3. In the screening with the synthetic oligonucleotide probe, on the other hand, the formamide concentration of the hybridization solution was adjusted to 20% and washing was repeated 5 times in a solution containing 0.3×SSC/0.1% of SDS at 45° C. each for 30 minutes.

With each probe, a library of 1,000,000 clones was screened. As a result, 2 positive plaques were obtained by using the probe NZ4 while 1 positive plaque was obtained by using the synthetic oligonucleotide probe. Each screening was repeated until all of the plaques became positive.

4) Extraction of λDNA

Each positive clone was propagated by using Escherichia coli LE392 as the host in accordance with the method described in Example 2 and then DNA was extracted.

(5) Partial Mapping and Sequencing

The λDNAs obtained in the above (4) (GHFc-1, 2 and 3) were completely digested with restriction enzymes (ApaI, BamHI, EcoRI, HindIII, KpnI, NcoI, PstI, SacI, ScaI, SmaI, SpeI, SphI, StuI, XbaI and XhoI), electrophoresed on a 1% agarose gel and then subjected to Southern blotting. Then each gel was immersed in 0.25 N HCl for 30 minutes and allowed to stand under shaking in a denaturation buffer (0.4 N NaOH/1.5 M NaCl) at room temperature for 15 minutes twice. Subsequently, it was further allowed to stand under shaking in a neutralization buffer (1 M NH₄OAc/0.02 N NaOH) at room temperature for 15 minutes twice. Next, each gel was transferred onto 2 nylon membrane (bidirectional transfer) and these 2 membranes were subjected to hybridization respectively with the probe NZ4 and the synthetic oligonucleotide probe.

Positive fragments of GHFc-1, 2 (ApaI, EcoRI, SacI and XhoI) and GHFc-3 (BamHI, EcoRI and XbaI) were subcloned into a pBluecript vector (manufactured by Toyobo) and subjected to partial sequencing. The sequencing was carried out in accordance with the method of Example 6.

(6) Result

As the results of the partial mapping and sequencing, it was found out that the clones GHFc-1, 2 and 3 were independent clones having inserts of about 15 kb (GHFC-1 and 2) and 13 kb (GHFc-3) and GHFc-1 partly overlapped GHFc-2. Introns satisfying the GT/AG rule were located between the bases at the 63- and 64-positions and those at the 1311- and 1312-positions of the cDNA of FcγBP (FIG. 10) and the base sequence of the exon region up to the 1311-position was completely identical with that of the cDNA (FIG. 11).

Further, the sequence located 5′ upstream of the cDNA clone NZ4 was contained in GHFc-3 and the stop codon (TGA) in frame was located 87 bases upstream from the assumed translation initiation codon ATG described in Example 7 (i.e., 79 bases upstream from the 5′ end of NZ4 represented by SEQ ID NO:1). No other ATG in frame was observed any more. These results strongly support the possibility that the ATG starting from the base at the 9-position in the clone NZ4 is the translation initiation ATG of the FcγBP gene.

Example 18 Correlation Between Structure of Human FcγBP and IgG Fc Region-binding Activity

The protein structure having the amino acid sequence assumed from the entire base sequence of the cDNA of FcγBP obtained so far is one having a unique sequence in which a unit consisting of about 400 amino acid residues is repeated 12 times in total (R1-R12 domain) and a sequence 25 consisting of about 450 amino acids (H domain) and another sequence consisting of about 200 amino acid (T domain) are located before and after the R1-R12 part. As FIG. 12 shows, among the repeating units in the R domain, the units R3, R6 and R9 have a homology of at least 95% with each other. Similarly, the units R4/R7/R10 and R5/R8/R11 ave each a homology of at least 95%. On the other hand, he units R1 to R5 have a homology of about 40% with each other. To discuss the function of these protein domains, mutated clones having some defects in the cDNA were separated and expressed in COS cells. Thus the functions including the activity of binding to IgG, etc. were examined.

As FIG. 12 shows, the partial cDNA plasmid pNV11, which had been expressed in animal cells in order to determine the Fc-binding activity, was composed of H, R1, R2, R3, R4, R5 and a part of R6. To clarify the unit having the Fc-biding activity, the following experiment was carried out. Namely, cDNA fragments, from which some part of NV11 had been deleted by cleaving and/or rebinding with restriction enzymes or ligating appropriate clones with each other so as to suit for an amino acid frame, were inserted into an expression vector SRα and then E. coli XL-1 was transformed thereby. Table 1 shows the sequences after the deletion of a part of NV11 expressed in the DNA base numbers. In the cDNA of NV11, the first base from which the translation into protein is initiated is referred to as No. 1 while the final base is referred to as No. 7776.

The expression vector containing each cDNA fragment was purified from Escherichia coli, tentatively expressed in COS7 cells in the same manner as the one of Example 11 and then stained with human IgG Fc region and the monoclonal antibodies K9 and K17 specific to FcγBP.

Further, the following staining was effected in order to examine the inhibition of the binding to the IgG Fc region by the monoclonal antibodies. The COS7 cells having cDNA tentatively expressed therein were fixed with ethanol. Then thermally denatured human IgG was diluted with hybridoma culture supernatants containing either or both of the inhibition antibodies K9 and K17 or antiFcγRIII (a control antibody) so as to give each a concentration of 1 μg/ml. Then the fixed cells were incubated together with each supernatant. After allowing to stand at room temperature for 1 hour and washing with PBS (−), the cells were further incubated together with HRP-labeled antihuman IgG (H+L) antibody F(ab7)₂ fragment and then the biding of the denatured IgG was detected.

The results are as follows. To examine whether or not the gene expression product had IgG Fc region-binding activity, human IgG was employed as the primary antibody while the HRP-labeled antihuman IgG antibody was employed as the secondary antibody. IgG-binding activity was observed in the clones having the entire sequence of the H domain and, furthermore, the entire region of at least one R unit (NV11, NX, NZCY, ΔBSSH, ΔTth, ΔSp1, ΔBssH/Tth, NXΔBssH and NZCV11). The strength of the stain indicating the IgG-binding activity was tend to increase with an increase in the number of R units contained in the clone. However, the clones from which the H domain had been completely or partly deleted (ΔHinc, ΔHinc/BssH, V11 and X1) and the clone having only a part of a R unit (NZC) showed no IgG-binding activity.

Regarding the staining of the gene products with the monoclonal antibodies, on the other hand, the clones having the entire or partial sequence of R5 (NV11, ΔHinc, ΔBssH, ΔTth, ΔSp1, ΔBssH/Tth, ΔHinc/BssH, NZCV11 and V11) were stained with the monoclonal antibody K9, while the clones having the entire or partial sequence of R3 or R6 (all clones except NZCY and NZC) were stained with the monoclonal antibody K17. These results indicated that in the clones lacking the H domain (ΔHinc/BssH, V11 and X1), no IgG-binding activity could be achieved, though the proteins reacting with the FcγBP-specific antibodies were produced therein. Thus it is suggested that the H domain would have a function essentially required in imparting the IgG-binding activity to the R domain products. The clone NZC showed no IgG-binding activity and was negative in the K9/K17 staining, which suggests that the R units (R1 to R5) might correspond to the IgG-binding site.

Subsequently, the results of the inhibition of the IgG-binding by the monoclonal antibodies K9 and K17 will be reported. Table 1 summarizes the inhibitory effects on each clone.

(1) The IgG-binding activities of the clones having at least one R unit in addition to R3 and R5 (NV11, ΔTth and NZCV11) were inhibited to a certain extent by K9 or K17. When these antibodies were added together, the inhibition was strengthened, though complete inhibition did not occur.

(2) The IgG-binding activity of the clone having at least one R unit in addition to R3 (NX) was inhibited by K17 not completely but to a certain extent. The activity of this clone was never inhibited by K9.

(3) The IgG-binding of the clone containing R3 alone (NXΔBssH) was completely inhibited by K17. In contrast, the binding was not affected by K9.

(4) The IgG-binding of the clone containing R5 alone (ΔBssH/Tth) was completely inhibited by K9. In contrast, the binding was not affected by K17.

(5) Both of K9 and K17 did not inhibit the IgG-binding of the clone containing neither R3 nor R5 (NZCY).

(6) The inhibition of the IgG-binding by the control antibody (antiFcγRIII antibody) was observed in none of the clones.

Based on these results, it is assumed that the IgG Fc binding sites are located in the R1-R5 region including R3 and R5 and each R unit can bind to IgG independently. Also, there is pointed out a possibility that 2 or more IgGs might be bound to cDNA clones having 2 or more R units (NV11, etc). These facts suggest that each of R1 to R12 might have the IgG binding site by taking the homology in the amino acid sequences into consideration.

TABLE 1 EXPRESSION TABLE Avidity to monoclonal IgG-binding antibody no Clone Base No. K9 K17 antibody +K9 +K17 +K9 + K17 NV11 1-7776 + + + + + + + + + + + + + + NX 1-4692 − + + + + + + + +   + NZCY 1-3013 − − + +   +   + NZC 1-1473 − − − ΔHinc 1-271, 1657-7776 + + + + + + − ΔBssH 1-1600, 2761-7776 + + + + + + + + ΔTth 1-3272, 5477-7776 + + + + + + + + + + + + + + ΔSp1 1-6066 + + + + + + + ΔBssH/Tth 1-1600, 2761-3272, 5477-7776 + + + + + + + +   − + + − NXΔBssH 1-1600, 2761-4692 − + + + +   + −   − ΔHinc/BssH 1-271, 2761-7776 + + + + + + − NZCV11 1-1473, 4165-7776 + + + + + + + + + + + + + + V11 4165-7776 − + + + − X1 1445-4692 − + + + −

Example 19 Partial Analysis of FcγBP Genome Gene (determination of transcription initiation site)

Based on the results of the primer extension carried out in Example 7, it was assumed that the transcription initiation site might be located within 20 bases upstream from the clone NZ4 which was located at the 5′-uppermost stream at the present stage; and that the ATG at the 9-position in NZ4 might be the translation initiation site. To examine whether or not ATG in frame or the stop codon was located upstream from NZ4, therefore, the FcγBP gene was isolated from the genomic DNA library and its partial base sequence was identified.

Further, the transcription initiation site was more accurately determined by S1 mapping.

(1) Genomic DNA Library

As the library, use was made of a commercially available one originating in human leukocytes (Vector, EMBL3 SP6/T7, manufactured by Clontech).

(2) Probe

As the probes to be employed in the screening, use was made of one prepared by excising the cDNA clone NZ4 from the vector with BamHI and the synthetic oligonucleotide employed in Example 7 (primer 2: (GCTCCAGCCCAGAGTATCCACCAGCTCCATAGG-33 mer, SEQ ID NO:11) labeled respectively with α[³²P] dCTP and γ[³²P] ATP by random prime labeling (NZ4) or end labeling (primer 2).

(3) Screening

The screening with the probe NZ4 was performed in accordance with the method for screening a cNDA library as described in Example 4. In the screening with the synthetic oligonucleotide probe, on the other hand, the formamide concentration of the hybridization solution was adjusted to 20% and washing was repeated 5 times in a solution containing 0.3×SSC/0.1% of SDS at 45° C. each for 30 minutes.

With each probe, a library of 1,000,000 clones was screened. As a result, 2 positive plaques were obtained by using the probe NZ4 while 1 positive plaque was obtained by using the synthetic oligonucleotide probe. Each screening was repeated until all of the plaques became positive.

(4) Extraction of λDNA

Each positive clone was propagated by using Escherichia coli LE392 as the host in accordance with the method described in Example 2 and then DNA was extracted.

(5) Partial Mapping and Sequencing

The λDNAs obtained in the above (4) (GHFc-1, 2 and 3) were completely digested with restriction enzymes (ApaI, BamHI, EcoRI, HindIII, KpnI, NcoI, PstI, SacI, ScaI, SmaI, SpeI, SphI, StuI, XbaI and XhoI), electrophoresed on a 1% agarose gel and then subjected to Southern blotting. Then each gel was treated with 0.25 N HCl, 0.4 N NaOH/1.5 M NaCl and 1 M NH₄Ac/0.02 N NaOH each for 15 minutes twice. Next, each gel was transferred onto 2 nylon membrane (bidirectional transfer) and these 2 membranes were subjected to hybridization respectively with the probe NZ4 and the synthetic oligonucleotide probe.

Positive fragments of GHFc-1, 2 (ApaI, EcoRI, SacI and XhoI) and GHFc-3 (BamHI, EcoRI and XhoI) were subcloned into a pBluescript vector (manufactured by Toyobo) and subjected to partial sequencing. The sequencing was carried out in accordance with the method of Example 6.

(6) S1 Mapping

As a template for constructing an S1 probe, use was made of ssDNA which had been prepared by subcloning the EcoRI/SacI fragment (corresponding to about 2 kb upstream from NZ4 in cDNA clone) of the clone GHFc-3 into pBluescript SK⁺ and treated with a helper phage VCSM13. To this template was annealed the labeled primer 2 employed in the primer extension followed by synthesis with BcaBEST polymerase (manufactured by Takara) at 65° C. for 10 minutes. Then the synthesis product was digested with BamHI, thermally denatured and separated by using a 7.5% polyacrylamide gel containing 8 M of urea. Then the target gel was excised. The gel thus excised was incubated in G buffer (1M of NH₄AC, 20 mM of Mg(OAc)₂, 0.1 M of EDTA, 0.2% of SDS, 10 μg/ml of yeast tRNA) at 37° C. overnight to thereby eluate the probe. This S1 probe (1×10⁵ cpm) was mixed with 40 μg of all RNAs originating inhuman colonic epithelial cells and 1.5 μg of polyA ⁺RNA. After precipitating from ethanol, the precipitate was dissolved in 20 μl of an S1 hybridization solution (80% of formamide, 40 mM of PIPES, 400 mM of NaCl, 1 mM of EDTA). After treating at 80° C. for 10 minutes, it was subjected to hybridization at 42° C. overnight. Then 200 μl of an S1 solution [30 mM of NaOAc (pH 4.6), 280 mM of NaCl, 1 mM of ZnSO₄, 1 mg/ml of ssDNA, 150 U of S1 nuclease] was added thereto followed by digestion at 37° C. for 40 minutes. Next, it was electrophoresed on a 6% sequence gel and the gel was fixed, dried and subjected to autoradiography.

RESULTS AND DISCUSSION

As the results of the partial mapping and sequencing, it was found out that the clones GHFc-1, 2 and 3 were independent clones having inserts of about 15 kb (GHFc-1 and 2) and 13 kb (GHFc-3) and GHFc-1 partly overlapped GHFc-2. Introns satisfying the GT/AG rule were located between the bases at the 63- and 64-positions and those at the 1311- and 1312-positions of the cDNA of FcγBP and the base sequence of the exon region up to the 1311-position was completely identical with that of the cDNA (FIGS. 10 and 11). Further, the sequence 5′ upstream from the cDNA clone NZ4 was contained in GHFc-3 and the stop codon (TGA) in frame was located 87 bases upstream from the assumed translation initiation codon ATG described in Example 7 (i.e., 79 bases upstream from the 5′ end of NZ4). No other ATG in frame was observed any more. These results strongly support the possibility that the ATG starting from the base at the 9-position in the clone NZ4 is the translation initiation ATG of the FcγBP gene. Further, no typical promoter motif (TATA/CCAAT, etc.) was contained within about 2 kbp 5′-upstream from NZ4. The results of the S1 mapping indicated that bands of the corresponding lengths were observed 8, 9 and 10 bases upstream from this ATG. Among these bands, the one of the A residue at the 10-position showed the strongest signal, which suggests that the transcription would be initiated from this A residue.

Example 20 Analysis of Polymorphism of FcγBP Gene

As the result of the sequencing of the cDNA of FcγBP, it was suggested that polymorphism occurred at specific sites in the coding region. Namely, the sequence CCCGGG at the SmaI sites located on the 5120-, 8723- and 12326-positions in the cDNA of FcγBP were replaced by CCTGGG in the same region of the clone A52. The library employed in the screening of the cDNA originated in genes of not a single person but several subjects. Accordingly, the following experiment was carried out in order to confirm whether the above-mentioned base replacement was observed among individuals or in the major repeating region (thrice) per haploid genome in a single individual.

The following 2 primers were synthesized as the primers on the forward side:

BC1: ACCACTCCTTCGATGGCC, SEQ ID NO:24, and

GS1: ACCTGTAACTATGTGCTGGC, SEQ ID NO:25.

On the other hand, the following 4 primers were synthesized as the primers on the reverse side:

GS2: TGGTGGTGACGGTGAAGGG, SEQ ID NO:26,

GS3: ACAGCAGGGTTGCCCCGG, SEQ ID NO:27,

GS4: TGGTGCCGAGGGCAGCCACG, SEQ ID NO:28, and

BC2: TGGGTCACTGAAATCCG, SEQ ID NO:29.

Further, leukocytes of 6 healthy subjects and colonic epithelial cells of normal parts of 4 patients with cancer were separated and DNAs were extracted therefrom by the method of Nelson et al. To 20 ng of each DNA was added primer sets BC1/BC2, BC1/GS3, BC1/GS4, GS1/GS3 and GS1/GS4 to give each a final concentration of 20 pmole. Then PCR was performed in a PCR buffer [10 mM of Tris-HCl (pH 8.3), 50 mM of KCl, 1.5 mM of MgCl₂, 200 μM of dNTPs, 0.001% of gelatin, 2.5 U of taq polymerase] in 30 cycles each cycle consisting of 94° C. for 1 minute, 60° C. for 1.5 minutes and 72° C. for 2.5 minutes. The PCR products were digested with SmaI, electrophoresed on a 2% agarose gel and then stained with EtBr.

As the results of the SmaI-treatment of the PCR products for the primer sets, it was found out that polymorphism was observed in each of the primer sets except BC1/BC2.

Namely, from among 10 DNA samples, one was completely digested with SmaI. Accordingly, this sample had the SmaI sites at last in all of the 6 repeating units involving the allele. In contrast, SmaI-digested products and undigested products were observed at various ratios in other samples. On the contrary, none of these 10 samples was free from any SmaI site. Furthermore, RT-PCR was performed in HT-29N2 cells by using the same primer sets followed by digestion with SmaI. As a result, all of the samples contained the sites.

The primers BC1/BC2 showed no polymorphism. This is seemingly because the PCR product had a chain length of about 1.8 kbp and thus there was an intron (about 1.6 kbp) between the primer GS4 and the primer BC2 and the samI site was located on the 5′ side of this intron. Thus it was assumed that no polymorphism was detected since the above-mentioned site was extremely close to the target SmaI site showing polymorphism and the BC1/BC2 amplification product was scarcely different in chain length from the SmaI-digestion product.

Example 21 Separation of High Expression-inducible CHO Cell Line and Detection of FcγBP

In order to establish a cell line capable of expressing a large mount of the FcγBP fragment described in Example 11 in a stable state, NV11ST (i.e., the partial FcγBP cDNA) was expressed by using an expression vector pMSXND for animal cells. The vector pMSXND is one having a metallothionein promoter, which can induce the expression of a protein with sodium butyrate, etc., as an expression promoter and being constructed in such a manner as to have dhfr gene which enables gene amplification after being integrated into chromosomal DNA.

(1) Alteration of Vector pMSXND

First, the plasmid was completely digested with XhoI corresponding to the cDNA cloning site of pMSXND. Subsequently, it was blunt-ended by treating with 0.4 U of Klenow fragment for 15 minutes.

Next, NotI linker (5′-pGCGGCCGC-3′) was ligated to the vector followed by complete digestion with NotI. After the completion of the autoligation, competent E. coli XL1-B was transformed thereby. The clones thus obtained were analyzed to thereby give a plasmid pMSXND-NOT in which the NotI linker had been inserted into the XhoI site of pMSXND.

(2) Insertion of cDNA

pMSND-NOT was completely digested with NotI and then dephosphorylated by treating with alkaline phosphatase. Next, the plasmid pNV11-ST, which was prepared in Example 6-(6) and had FcγBP cDNA integrated thereinto, was completely digested with NotI and electrophoresed on an agarose gel to thereby separate and recover a cDNA fragment of 8 kbp. These expression vectors were ligated to the cDNA insert and competent E. coli (XL1-B) was transformed thereby. From the colonies formed by incubating the transformant on an LB plate containing ampicillin, a plasmid in which cDNA had been inserted in the sense direction to the metallothionein promoter was selected and referred to as pNV11-MSX.

(3) Expression of Partial cDNA of IgG FcBP Region in CHO Cell

Escherichia coli containing the plasmid pNV11-MSX was incubated in the same manner as the one described in Example 11 and the plasmid thus amplified was purified. Then 10 μg of this plasmid dissolved in 250 μl of F-12 medium (containing nucleotide) was mixed with 10 μl of a lipofection reagent (Transfectum, manufactured by Sepacor) dissolved in 250 μl of F-12 medium and the resulting mixture was immediately added onto CHO cells (dhfr-deficient line). After incubating at 37° C. for 6 hours, the medium was replaced by F-12 medium containing 10% of serum and the incubation was continued for 3 days. Next, the medium was replaced by α-MEM medium (nucleotide-free, manufactured by Gibco) containing 1 mg/ml of G418 and 10% of fetal bovine serum. Then incubation was carried out for 14 days while replacing the medium at intervals of 3 days. Next, cells having the plasmid inserted thereinto were selected. Thus the cells were cloned by the limiting dilution analysis from a dish in which several ten colonies had been formed. Among the cell clones thus obtained, those showing the expression of the protein with high Fc-binding activity were selected. Subsequently, gene amplification was carried out in order to elevate the expression yield.

(4) Gene Amplification

The integrated gene was amplified by treating with methotrexate so as to elevate the expression yield of the protein by the cell clone which had pNV11-MSX integrated into the chromosome of CHO cells and was capable of stably expressing the FcγBP fragment.

Namely, the above-mentioned cell clone showing stable expression was incubated in A-MEM medium (nucleotide-free) containing 0.005 μM of methotrexate and 500 μg/ml of G418 for 3 to 4 weeks and the cells thus grown were selected. Next, the methotrexate concentration was elevated 4-fold (0.02 μM) and the incubation was continued in the same manner for additional 3 to 4 weeks. The procedure of elevating the methotrexate concentration 4-fold followed by incubation was repeated to thereby finally give cells propagated in the presence of 6.4 to 25 μM of methotrexate. These cells were cloned by the limiting dilution analysis to thereby give a cell line with high expression of the FcγBP fragment. Similar to Examples 11 and 12, the increase in the expression yield was determined by the primary judgement by histochemical staining with the monoclonal antibodies K9/K17 or the detection of the IgG-binding activity.

(5) Preparation of Cell Sample

5×10⁵ cells of the CHO cell line, which could stably express the FcγBP fragment at a high yield, were transferred into a dish (diameter: 100 mm) and incubated in α-MEM medium containing 6.4 μM of methotrexate and 1 mg/ml of G418. If necessary, sodium butyrate was added in such an amount as to give a final concentration of 5 mM to thereby induce the expression of the protein. After 3 days, the culture supernatant (SUPL) was recovered. Then the SUP1 was centrifuged at 100,000×g for 60 minutes to thereby eliminate cell pieces therefrom and the supernatant (SUP2) was recovered. The cells were suspended in 500 μl of a cytolysis buffer [50 mM of Tris-HCl (pH 7.5), 150 mM of NaCl, 1 mM of EDTA, 1 mM of PMSF, 10 mM of monoiodo acetamide, 10 μg/ml of approtinine, 10 μg/ml of leupeptin] and subjected to ultrasonication (30 seconds×3 times) and centrifugation at 10,000×g for 10 minutes at 4° C. After the completion of the centrifugation, the supernatant (LYS1) was eliminated. To the residue was added 400 μl of the cytolysis buffer containing 1% of NP-40 followed by ultrasonication and centrifugation. After recovering the supernatant (LYS2), the residue was dissolved in 200 μl of the cytolysis buffer containing 1% of NP-40, 0.1% of SDS and 0.5% of sodium deoxycholate followed by ultrasonication and centrifugation. After recovering the supernatant (LYS3), the residue was dissolved in 100 μl of the cytolysis buffer (LYS4). As a control, use was made of solutions of colonic epithelial cells (diluted 100- to 3,200-fold).

(6) Sandwich ELISA

To quantitatively detect the FcγBP fragment thus produced, a sandwich ELISA system with the use of the monoclonal antibodies K9 and K17 specific for FcγBP was developed. The antibody K9 purified by affinity chromatography was dissolved in a 0.05 M carbonate buffer (pH 9.2) in such a manner as to give a concentration of 5 μg/ml and added to an ELISA plate (PRO-BIND, manufactured by Falcon) at a ratio of 50 μl/well. After allowing to stand at 4° C. overnight, the wells were washed with a washing liquor [PBS (−) containing 0.05% of Tween-20] thrice. Then 50 μl/well of a blocking solution (RPMI 1640 medium containing 10% of serum) was added and the plate was allowed to stand at room temperature for 60 minutes.

After eliminating the blocking solution, 50 μl portions of the samples prepared in the above (5) were added and the plate was allowed to stand at room temperature for 2 hours. After washing with the washing liquor thrice, 50 μl of HRP-labeled K17 antibody, which had been diluted with the blocking solution to give a concentration of 4 mg/ml, was added and the plate was allowed to stand at room temperature for 1 hour. Then it was washed with the washing liquor thrice and 50 μl of a color development solution [20 mg of o-phenylenediamine and 80 μl of H₂O₂ (30%) dissolved in 50 ml of citrate buffer] was added thereto followed by the color development at room temperature for 3 minutes. Subsequently, the reaction was ceased by adding 50 μl of a 2.5 M H₂SO₄ solution and the absorbance at 492 nm was measured.

RESULT

(i) Typical CHO Cell Line Showing FcγBP Expression Obtained Above

After incubating in the presence of methotrexate optionally followed by the treatment with sodium butyrate, the expression yield of the FcγBP fragment after 3 days was detected by the cell staining method with the use of the monoclonal antibodies K9/K17. Thus a cell line with a high expression yield could be isolated as shown in FIG. 13.

(ii) Result of Determination of FcγBP Fragment by ELISA

The samples obtained from the cell line with a high expression yield of the FcγBP fragment were subjected to sandwich ELISA. As a result, the cytolysis solutions (LYS1 to LYS4) contained the FcγBP fragment in larger amounts than the culture supernatants (SUP1 and SUP2), as shown in the absorbances listed in the following Table 2. This fact indicates that the FcγBP fragment thus expressed is not accumulated in the cells but secreted out therefrom.

TABLE 2 Sample OD₄₉₂ colonic epithelial cell lysate dilution (control) 100-fold 2.184 200 2.024 400 1.516 800 0.935 1600 0.514 3200 0.264 expression-induced cell SUP1 0.731 SUP2 0.712 LYS1 0.044 LYS2 0.174 LYS3 0.013 LYS4 0.095 expression-uninduced cell SUP1 0.259 SUP2 0.273 LYS1 0.027 LYS2 0.070 LYS3 0.012 LYS4 0.019

29 908 base pairs nucleic acid double linear cDNA not provided 1 CTGCAGCCAT GGGTGCCCTA TGGAGCTGGT GGATACTCTG GGCTGGAGCA ACCCTCCTGT 60 GGGGATTGAC CCAGGAGGCT TCAGTGGACC TCAAGAACAC TGGCAGAGAG GAATTCCTCA 120 CAGCCTTCCT GCAGAACTAT CAGCTGGCCT ACAGCAAGGC CTACCCCCGC CTCCTTATCT 180 CCAGTCTGTC AGAGAGCCCC GCTTCAGTCT CCATCCTCAG CCAGGCAGAC AACACCTCAA 240 AGAAGGTCAC AGTGAGGCCC GGGGAGTCGG TCATGGTCAA CATCAGTGCC AAGGCTGAGA 300 TGATAGGCAG CAAGATCTTC CAGCATGCGG TGGTGATCCA TTCTGACTAT GCCATCTCTG 360 TGCAGGCACT AAATGCCAAG CCTGACACAG CGGAGCTGAC ACTGCTGCGG CCCATCCAGG 420 CCCTAGGCAC CGAGTATTTT GTGCTCACAC CCCCCGGCAC CTCAGCCAGG AATGTCAAGG 480 AGTTTGCCGT GGTGGCCGGT GCCGCAGGTG CCTCGGTCAG TGTCACGCTG AAGGGGTCAG 540 TGACATTCAA TGGCAAGTTC TATCCAGCAG GCGATGTCCT AAGAGTGACT CTACAGCCCT 600 ACAATGTGGC CCAGCTACAG AGCTCAGTGG ATCTCTCGGG GTCAAAGGTC ACAGCTAGTA 660 GCCCCGTGGC TGTCCTCTCT GGCCACAGCT GTGCGCAGAA ACATACGACC TGCAACCATG 720 TGGTTGAGCA GCTGCTACCC ACGTCTGCCT GGGGCACCCA CTATGTAGTA CCCACGCTGG 780 CCTCCCAATC TCGCTATGAT TTGGCCTTCG TTGTGGCCAG CCAGGCCACA AAGCTGACCT 840 ACAACCATGG GGGTATCACT GGCTCCCGTG GGCTCCAGGC AGGTGATGTG GTAGAGTTTG 900 AGGTCCGG 908 1336 base pairs nucleic acid double linear cDNA not provided 2 GGCCTACAGC AAGGCCTACC CCCGCCTCCT TATCTCCAGT CTGTCAGAGA GCCCCGCTTC 60 AGTCTCCATC CTCAGCCAGG CAGACAACAC CTCAAAGAAG GTCACAGTGA GGCCCGGGGA 120 GTCGGTCATG GTCAACATCA GTGCCAAGGC TGAGATGATA GGCAGCAAGA TCTTCCAGCA 180 TGCGGTGGTG ATCCATTCTG ACTATGCCAT CTCTGTGCAG GCACTAAATG CCAAGCCTGA 240 CACAGCGGAG CTGACACTGC TGCGGCCCAT CCAGGCCCTA GGCACCGAGT ATTTTGTGCT 300 CACACCCCCC GGCACCTCAG CCAGGAATGT CAAGGAGTTT GCCGTGGTGG CCGGTGCCGC 360 AGGTGCCTCG GTCAGTGTCA CGCTGAAGGG GTCAGTGACA TTCAATGGCA AGTTCTATCC 420 AGCAGGCGAT GTCCTAAGAG TGACTCTACA GCCCTACAAT GTGGCCCAGC TACAGAGCTC 480 AGTGGATCTC TCGGGGTCAA AGGTCACAGC TAGTAGCCCC GTGGCTGTCC TCTCTGGCCA 540 CAGCTGTGCG CAGAAACATA CGACCTGCAA CCATGTGGTT GAGCAGCTGC TACCCACGTC 600 TGCCTGGGGC ACCCACTATG TAGTACCCAC GCTGGCCTCC CAATCTCGCT ATGATTTGGC 660 CTTCGTTGTG GCCAGCCAGG CCACAAAGCT GACCTACAAC CATGGGGGTA TCACTGGCTC 720 CCGTGGGCTC CAGGCAGGTG ATGTGGTAGA GTTTGAGGTC CGGCCATCCT GGCCACTCTA 780 CCTGTCTGCA AATGTGGGCA TCCAGGTCCT GTTGTTTGGC ACAGGTGCCA TAAGGAATGA 840 AGTGACTTAT GACCCCTACC TGGTCCTGAT CCCAGATGTG GCGGCCTACT GCCCAGCCTA 900 TGTGGTCAAG AGTGTACCAG GCTGTGAGGG CGTGGCCCTG GTAGTGGCAC AGACGAAGGC 960 TATCAGCGGG CTGACCATAG ATGGGCATGC AGTGGGGGCC AAGCTCACCT GGGAGGCTGT 1020 GCCAGGCAGT GAGTTCTCGT ATGCTGAAGT GGAGCTCGGC ACAGCTGACA TGATCCACAC 1080 GGCCGAGGCC ACCACCAACT TGGGACTGCT CACCTTCGGG CTGGCCAAGG CTATAGGCTA 1140 CGCAACAGCT GCTGATTGCG GCCGGACTGT ACTGTCCCCA GTGGAGCCCT CCTGCGAAGG 1200 CATGCAGTGC GCAGCCGGGC AGCGCTGCCA GGTGGTAGGC GGGAAGGCCG GGTGTGTGGC 1260 GGAGTCCACC GCTGTCTGCC GCGCCCAGGG CGACCCCCAT TACACCACCT TCGACGGCCG 1320 TCGCTACGAC ATGATG 1336 1878 base pairs nucleic acid double linear cDNA not provided 3 CCAAGCTCAC CTGGGAGGCT GTGCCAGGCA GTGAGTTCTC GTATGCTGAA GTGGAGCTCG 60 GCACAGCTGA CATGATCCAC ACGGCCGAGG CCACCACCAA CTTGGGACTG CTCACCTTCG 120 GGCTGGCCAA GGCTATAGGC TACGCAACAG CTGCTGATTG CGGCCGGACT GTACTGTCCC 180 CAGTGGAGCC CTCCTGCGAA GGCATGCAGT GCGCAGCCGG GCAGCGCTGC CAGGTGGTAG 240 GCGGGAAGGC CGGGTGTGTG GCGGAGTCCA CCGCTGTCTG CCGCGCCCAG GGCGACCCCC 300 ATTACACCAC CTTCGACGGC CGTCGCTACG ACATGATGGG CACCTGTTCG TACACGATGG 360 TGGAGCTGTG CAGCGAGGAC GACACCCTGC CCGCCTTCAG CGTGGAGGCC AAGAACGAGC 420 ACCGGGGCAG CCGCCGCGTC TCCTACGTGG GCCTCGTCAC TGTGCGCGCC TACAGCCACT 480 CTGTGTCGCT GACCCGCGGT GAAGTTGGCT TCGTCCTGGT TGACAACCAG CGCTCGCGCC 540 TGCCAGTCTC CCTGAGTGAG GGTCGCCTGC GTGTGTACCA GAGCGGACCA CGGGCCGTGG 600 TGGAGCTGGT CTTTGGGCTG GTGGTCACTT ATGACTGGGA CTGCCAGCTG GCACTCAGCC 660 TGCCTGCACG CTTCCAAGAC CAGGTGTGCG GGCTGTGTGG CAACTATAAT GGTGACCCAG 720 CAGACGACTT CCTCACGCCT GACGGGGCTC TGGCTCCTGA CGCTGTGGAG TTCGCAAGTA 780 GCTGGAAGCT GGATGATGGG GACTACCTGT GTGAGGATGG CTGCCAGAAC AACTGTCCCG 840 CCTGCACCCC AGGCCAGGCC CAACACTATG AGGGCGACCG ACTCTGTGGC ATGCTGACCA 900 AGCTCGATGG CCCCTTCGCT GTCTGCCATG ACACCCTGGA CCCCAGGCCC TTCCTGGAGC 960 AGTGTGTATA TGACCTGTGT GTGGTCGGTG GGGAGCGGCT CAGCCTGTGC CGTGGCCTCA 1020 GCGCCTATGC CCAGGCCTGT CTGGAGCTTG GCATCTCGGT TGGGGACTGG AGATCACCAG 1080 CCAACTGCCC CCTGTCCTGC CCTGCCAACA GCCGCTATGA GCTCTGCGGC CCTGCTTGCC 1140 CGACCTCCTG CAACGGGGCT GCGGCGCCGT CCAACTGCTC CGGGCGCCCC TGCGTGGAGG 1200 GCTGCGTGTG CCTCCCAGGC TTCGTGGCCA GCGGCGGCGC CTGCGTGCCG GCCTCGTCGT 1260 GTGGCTGCAC CTTCCAGGGT CTCCAGCTCG CTCCGGGCCA GGAAGTGTGG GCGGACGAGT 1320 TGTGCCAAAG GCGCTGCACC TGCAACGGCG CCACCCATCA GGTCACCTGC CGCGACAAGC 1380 AGAGCTGCCC GGCGGGTGAG CGCTGCAGCG TCCAGAACGG CCTCCTGGGC TGCTACCCCG 1440 ATCGCTTCGG GACCTGCCAG GGGTCCGGGG ACCCACACTA TGTGAGCTTC GACGGCCGGC 1500 GCTTCGACTT CATGGGCACC TGCACGTACC TGCTGGTCGG CTCATGCGGC CAGAACGCAG 1560 CGCTGCCTGC CTTCCGGGTG CTGGTGGAAA ACGAGCATCG GGGCAGCCAG ACTGTGAGCT 1620 ACACGCGCGC CGTGCGGGTG GAGGCCCGCG GGGTGAAGGT GGCCGTGCGC CGGGAGTACC 1680 CCGGGCAAGT GCTGGTGGAT GACGTCCTTC AGTATCTGCC CTTCCAAGCA GCAGATGGGC 1740 AGGTGCAGGT GTTCCGACAG GGCAGGGATG CCGTCGTGCG CACGGACTTT GGCCTGACTG 1800 TCACTTATGA CTGGAATGCA CGAGTGACTG CCAAGGTGCC CAGCAGCTAT GCTGAGGCCC 1860 TGTGTGGACT CTGTGGGA 1878 3247 base pairs nucleic acid double linear cDNA not provided 4 CTTCGACGGC CGTCGCTACG ACATGATGGG CACCTGTTCG TACACGATGG TGGAGCTGTG 60 CAGCGAGGAC GACACCCTGC CCGCCTTCAG CGTGGAGGCC AAGAACGAGC ACCGGGGCAG 120 CCGCCGCGTC TCCTACGTGG GCCTCGTCAC TGTGCGCGCC TACAGCCACT CTGTGTCGCT 180 GACCCGCGGT GAAGTTGGCT TCGTCCTGGT TGACAACCAG CGCTCGCGCC TGCCAGTCTC 240 CCTGAGTGAG GGTCGCCTGC GTGTGTACCA GAGCGGACCA CGGGCCGTGG TGGAGCTGGT 300 CTTTGGGCTG GTGGTCACTT ATGACTGGGA CTGCCAGCTG GCACTCAGCC TGCCTGCACG 360 CTTCCAAGAC CAGGTGTGCG GGCTGTGTGG CAACTATAAT GGTGACCCAG CAGACGACTT 420 CCTCACGCCT GACGGGGCTC TGGCTCCTGA CGCTGTGGAG TTCGCAAGTA GCTGGAAGCT 480 GGATGATGGG GACTACCTGT GTGAGGATGG CTGCCAGAAC AACTGTCCCG CCTGCACCCC 540 AGGCCAGGCC CAACACTATG AGGGCGACCG ACTCTGTGGC ATGCTGACCA AGCTCGATGG 600 CCCCTTCGCT GTCTGCCATG ACACCCTGGA CCCCAGGCCC TTCCTGGAGC AGTGTGTATA 660 TGACCTGTGT GTGGTCGGTG GGGAGCGGCT CAGCCTGTGC CGTGGCCTCA GCGCCTATGC 720 CCAGGCCTGT CTGGAGCTTG GCATCTCGGT TGGGGACTGG AGATCACCAG CCAACTGCCC 780 CCTGTCCTGC CCTGCCAACA GCCGCTATGA GCTCTGCGGC CCTGCTTGCC CGACCTCCTG 840 CAACGGGGCT GCGGCGCCGT CCAACTGCTC CGGGCGCCCC TGCGTGGAGG GCTGCGTGTG 900 CCTCCCAGGC TTCGTGGCCA GCGGCGGCGC CTGCGTGCCG GCCTCGTCGT GTGGCTGCAC 960 CTTCCAGGGT CTCCAGCTCG CTCCGGGCCA GGAAGTGTGG GCGGACGAGT TGTGCCAAAG 1020 GCGCTGCACC TGCAACGGCG CCACCCATCA GGTCACCTGC CGCGACAAGC AGAGCTGCCC 1080 GGCGGGTGAG CGCTGCAGCG TCCAGAACGG CCTCCTGGGC TGCTACCCCG ATCGCTTCGG 1140 GACCTGCCAG GGGTCCGGGG ACCCACACTA TGTGAGCTTC GACGGCCGGC GCTTCGACTT 1200 CATGGGCACC TGCACGTACC TGCTGGTCGG CTCATGCGGC CAGAACGCAG CGCTGCCTGC 1260 CTTCCGGGTG CTGGTGGAAA ACGAGCATCG GGGCAGCCAG ACTGTGAGCT ACACGCGCGC 1320 CGTGCGGGTG GAGGCCCGCG GGGTGAAGGT GGCCGTGCGC CGGGAGTACC CCGGGCAAGT 1380 GCTGGTGGAT GACGTCCTTC AGTATCTGCC CTTCCAAGCA GCAGATGGGC AGGTGCAGGT 1440 GTTCCGACAG GGCAGGGATG CCGTCGTGCG CACGGACTTT GGCCTGACTG TCACTTATGA 1500 CTGGAATGCA CGAGTGACTG CCAAGGTGCC CAGCAGCTAT GCTGAGGCCC TGTGTGGACT 1560 CTGTGGGAAC TTCAACGGGG ACCCAGCTGA TGACCTGGCT CTGCGGGGTG GGGGTCAAGC 1620 TGCCAATGCA CTGGCCTTTG GGAACAGCTG GCAAGAAGAG ACGAGGCCCG GCTGTGGAGC 1680 AACTGAACCG GGTGACTGTC CCAAGCTGGA CTCCCTGGTG GCCCAGCAGC TGCAGAGCAA 1740 GAATGAGTGT GGAATCCTTG CCGACCCCAA GGGGCCCTTC CGGGAGTGCC ATAGCAAGCT 1800 GGACCCCCAG GGTGCCGTGC GCGACTGTGT CTATGACCGC TGCCTGCTGC CAGGCCAGTC 1860 TGGGCCACTG TGTGACGCAC TGGCCACCTA TGCTGCTGCA TGCCAGGCTG CTGGAGCCAC 1920 AGTGCACCCC TGGAGGAGTG AAGAACTTTG CCCACTGAGC TGCCCACCCC ACAGCCACTA 1980 TGAGGCGTGT TCCTACGGCT GCCCGCTGTC CTGTGGAGAC CTCCCAGTGC CCGGGGGCTG 2040 TGGCTCAGAA TGCCATGAGG GCTGCGTGTG CGATGAGGGC TTTGCGCTCA GTGGTGAGTC 2100 CTGCCTGCCC CTGGCCTCCT GTGGCTGCGT ACACCAGGGC ACCTACCACC CACCAGGCCA 2160 GACCTTCTAC CCTGGCCCCG GATGTGATTC CCTTTGCCAC TGCCAGGAGG GCGGCCTGGT 2220 GTCCTGTGAG TCCTCCAGCT GCGGACCGCA CGAGGCCTGC CAGCCATCCG GTGGCAGCTT 2280 GGGCTGTGTG GCCGTGGGCT CTAGCACCTG CCAGGCGTCA GGAGACCCCC ACTACACCAC 2340 CTTCGATGGC CGCCGCTTCG ACTTCATGGG CACCTGCGTG TATGTGCTGG CTCAGACCTG 2400 CGGCACCCGG CCTGGCCTGC ATCGGTTTGC CGTCCTGCAG GAGAACGTGG CCTGGGGTAA 2460 TGGGCGAGTC AGTGTGACCA GGGTGATCAC GGTCCAGGTG GCAAACTTCA CCCTGCGGCT 2520 GGAGCAGAGA CAGTGGAAGG TCACGGTGAA CGGTGTGGAC ATGAAGCTGC CCGTGGTGCT 2580 GGCCAACGGC CAGATCCGTG CCTCCCAGCA TGGTTCAGAT GTTGTGATTG AGACCGACTT 2640 CGGCCTGCGT GTGGCCTACG ACCTTGTGTA CTATGTGCGG GTCACCGTCC CCGGAAACTA 2700 CTACCAGCAG ATGTGTGGCC TGTGTGGGAA CTACAACGGC GACCCCAAGG ATGACTTCCA 2760 GAAGCCCAAT GGCTCACAGG CAGGCAACGC CAATGAGTTC GGCAACTCCT GGGAGGAGGT 2820 GGTGCCCGAC TCTCCCTGCC TGCCGCCCAC CCCTTGCCCG CCGGGGAGCG AGGACTGTAT 2880 CCCCAGCCAC AAGTGTCCTC CCGAGCTGGA GAAGAAGTAT CAGAAGGAGG AGTTCTGTGG 2940 GCTCCTCTCC AGCCCCACAG GGCCACTGTC CTCCTGCCAC AAGCTGGTGG ATCCCCAGGG 3000 TCCCTTGAAA GATTGCATCT TTGATCTCTG CCTGGGTGGT GGGAACCTGA GCATTCTCTG 3060 CAGCAACATC CATGCCTACG TGAGTGCTTG CCAGGCGGCT GGAGGCCACG TGGAGCCCTG 3120 GAGGACTGAA ACTTTCTGTC CCATGGAGTG CCCTCCGAAC AGTCACTACG AGCTCTGTGC 3180 GGACACCTGC TCCCTGGGCT GCTCAGCTCT CAGTGCCCCT CCACAGTGCC AGGATGGGTG 3240 TGCTGAG 3247 3661 base pairs nucleic acid single linear cDNA not provided 5 CTATGTGCGG GTCACCGTCC CCGGAAACTA CTACCAGCAG ATGTGTGGCC TGTGTGGGAA 60 CTACAACGGC GACCCCAAGG ATGACTTCCA GAAGCCCAAT GGCTCACAGG CAGGCAACGC 120 CAATGAGTTC GGCAACTCCT GGGAGGAGGT GGTGCCCGAC TCTCCCTGCC TGCCGCCCAC 180 CCCTTGCCCG CCGGGGAGCG AGGACTGTAT CCCCAGCCAC AAGTGTCCTC CCGAGCTGGA 240 GAAGAAGTAT CAGAAGGAGG AGTTCTGTGG GCTCCTCTCC AGCCCCACAG GGCCACTGTC 300 CTCCTGCCAC AAGCTGGTGG ATCCCCAGGG TCCCTTGAAA GATTGCATCT TTGATCTCTG 360 CCTGGGTGGT GGGAACCTGA GCATTCTCTG CAGCAACATC CATGCCTACG TGAGTGCTTG 420 CCAGGCGGCT GGAGGCCACG TGGAGCCCTG GAGGACTGAA ACTTTCTGTC CCATGGAGTG 480 CCCTCCGAAC AGTCACTACG AGCTCTGTGC GGACACCTGC TCCCTGGGCT GCTCAGCTCT 540 CAGTGCCCCT CCACAGTGCC AGGATGGGTG TGCTGAGGGC TGCCAGTGTG ACTCCGGCTT 600 CCTCTACAAT GGCCAAGCCT GCGTGCCCAT CCAGCAATGC GGCTGCTACC ACAATGGTGT 660 CTACTATGAG CCGGAGCAGA CAGTCCTCAT TGACAACTGT CGGCAGCAGT GCACGTGCCA 720 TGCGGGTAAA GGCATGGTGT GCCAGGAACA CAGCTGCAAG CCGGGGCAGG TGTGCCAGCC 780 CTCCGGAGGC ATCCTGAGCT GCGTCACCAA AGACCCGTGC CACGGCGTGA CATGCCGGCC 840 ACAGGAGACA TGCAAGGAGC AGGGTGGCCA GGGCGTGTGC CTGCCCAACT ATGAGGCCAC 900 GTGCTGGCTG TGGGGCGACC CACACTACCA CTCCTTCGAT GGCCGGAAGT TTGACTTCCA 960 GGGCACCTGT AACTATGTGC TGGCAACAAC TGGCTGCCCG GGGGTCAGCA CCCAGGGCCT 1020 GACACCCTTC ACCGTCACCA CCAAGAACCA GAACCGGGGC AACCCTGCTG TGTCCTACGT 1080 GAGAGTCGTC ACCGTGGCTG CCCTCGGCAC CAACATCTCC ATCCACAAGG ACGAGATCGG 1140 CAAAGTCCGG GTGAACGGTG TGCTCACAGC CTTGCCTGTC TCTGTGGCCG ACGGGCGGAT 1200 TTCAGTGACC CAGGGTGCAT CGAAGGCACT GCTGGTGGCT GACTTTGGAC TGCAAGTCAG 1260 CTATGACTGG AACTGGCGGG TAGACGTGAC GCTGCCCAGC AGCTATCATG GCGCAGTGTG 1320 CGGGCTCTGC GGTAACATGG ACCGCAACCC CAACAATGAC CAGGTCTTCC CTAATGGCAC 1380 ACTGGCTCCC TCCATACCCA TCTGGGGCGG CAGCTGGCGA GCCCCAGGCT GGGACCCACT 1440 GTGTTGGGAC GAATGTCGGG GGTCCTGCCC AACGTGCCCT GAGGACCGGT TGGAGCAGTA 1500 CGAGGGCCCT GGCTTCTGCG GACCCCTGGC CCCCGGCACA GGGGGCCCTT TCACCACCTG 1560 CCATGCTCAT GTGCCACCTG AGAGCTTCTT CAAGGGCTGT GTTCTGGACG TCTGCATGGG 1620 TGGTGGGGAC CGTGACATTC TTTGCAAGGC TCTGGCTTCC TATGTGGCCG CCTGCCAGGC 1680 TGCTGGGGTT GTCATCGAAG ACTGGCGGGC ACAGGTTGGC TGTGAGATCA CCTGCCCAGA 1740 AAACAGCCAC TATGAGGTCT GTGGCCCACC CTGCCCGGCC AGCTGTCCGT CCCCTGCACC 1800 CCTTACGACG CCAGCCGTAT GTGAGGGCCC CTGTGTGGAG GGCTGCCAGT GCGACGCGGG 1860 TTTCGTGTTA AGTGCTGACC GCTGTGTTCC CCTCAACAAC GGCTGCGGCT GCTGGGCCAA 1920 TGGCACCTAC CACGAGGCGG GCAGTGAGTT TTGGGCTGAT GGCACCTGCT CCCAGTGGTG 1980 TCGCTGCGGG CCTGGGGGTG GCTCGCTGGT CTGCACACCT GCCAGCTGTG GGCTGGGTGA 2040 AGTGTGTGGC CTCCTGCCAT CCGGCCAGCA CGGCTGCCAG CCCGTCAGCA CAGCTGAGTG 2100 CCAGGCGTGG GGTGACCCCC ATTACGTCAC TCTGGATGGG CACCGATTCA ATTTCCAAGG 2160 CACCTGCGAG TACCTGCTGA GTGCACCCTG CCACGGACCA CCCTTGGGGG CTGAGAACTT 2220 CACTGTCACT GTAGCCAATG AGCACCGGGG CAGCCAGGCT GTCAGCTACA CCCGCAGTGT 2280 CACCCTGCAA ATCTACAACC ACAGCCTGAC ACTGAGTGCC CGCTGGCCCC GGAAGCTACA 2340 GGTGGACGGC GTGTTCGTCA CTCTGCCCTT CCAGCTGGAC TCGCTCCTGC ACGCACACCT 2400 GAGCGGCGCC GACGTGGTGG TGACCACAAC CTCAGGGCTC TCGCTGGCTT TCGACGGGGA 2460 CAGCTTCGTG CGCCTGCGCG TGCCGGCGGC GTACGCGGGC TCTCTCTGTG GCTTATGCGG 2520 GAACTACAAC CAGGACCCCG CAGACGACCT GAAGGCGGTG GGCGGGAAGC CCGCCGGATG 2580 GCAGGTGGGC GGCGCCCAGG GCTGCGGGGA ATGTGTGTCC AAGCCATGCC CGTCGCCGTG 2640 CACCCCAGAG CAGCAAGAGT CCTTCGGCGG CCCGGACGCC TGCGGCGTGA TCTCCGCCAC 2700 CGACGGCCCG CTGGCGCCCT GCCACGGCCT TGTGCCGCCC GCGCAGTACT TCCAGGGCTG 2760 CTTGCTGGAC GCCTGCCAAG TTCAGGGCCA TCCTGGAGGC CTCTGTCCTG CAGTGGCCAC 2820 CTACGTGGCA GCCTGTCAGG CCGCTGGGGC CCAGCTCCGC GAGTGGAGGC GGCCGGACTT 2880 CTGTCCCTTC CAGTGCCCTG CCCACAGCCA CTACGAGCTC TGCGGTGACT CCTGTCCTGG 2940 GAGCTGCCCG AGCCTGTCGG CACCCGAGGG CTGTGAGTCG GCCTGCCGTG AAGGCTGTGT 3000 CTGCGATGCT GGCTTCGTGC TCAGTGGTGA CACGTGTGTA CCTGTGGGCC AGTGTGGCTG 3060 CCTCCACGAT GACCGCTACT ACCCACTGGG CCAGACCTTC TACCCTGGCC CTGGGTGTGA 3120 TTCCCTTTGC CGCTGCCGGG AGGGCGGTGA GGTGTCCTGT GAGCCCTCCA GCTGCGGCCC 3180 GCATGAGACC TGCCGGCCAT CCGGTGGCAG CTTGGGCTGC GTGGCCGTGG GCTCTACCAC 3240 CTGCCAGGCG TCGGGAGATC CCCACTACAC CACCTTCGAT GGCCGCCGCT TCGACTTCAT 3300 GGGCACCTGC GTGTATGTGC TGGCTCAGAC CTGCGGCACC CGGCCTGGCC TACATCGGTT 3360 TGCCGTCCTG CAGGAGAACG TGGCCTGGGG TAATGGGCGA GTCAGTGTGA CCAGGGTGAT 3420 CACGGTCCAG GTGGCAAACT TCACCCTGCG GCTGGAGCAG AGACAGTGGA AGGTCACGGT 3480 GAACGGTGTG GACATGAAGC TGCCCGTGGT GCTGGCCAAC GGCCAGATCC GTGCCTCCCA 3540 GCATGGTTCA GATGTTGTGA TTGAGACCGA CTTCGGCCTG CGTGTGGCCT ACGACCTTGT 3600 GTACTATGTG CGGGTCACCG TCCCTGGAAA CTACTACCAG CTGATGTGTG GCCTGTGTGG 3660 G 3661 7824 base pairs nucleic acid double linear cDNA not provided CDS 21..7802 6 GGCCGCGGAT CCCTGCAGCC ATG GGT GCC CTA TGG AGC TGG TGG ATA CTC 50 Met Gly Ala Leu Trp Ser Trp Trp Ile Leu 1 5 10 TGG GCT GGA GCA ACC CTC CTG TGG GGA TTG ACC CAG GAG GCT TCA GTG 98 Trp Ala Gly Ala Thr Leu Leu Trp Gly Leu Thr Gln Glu Ala Ser Val 15 20 25 GAC CTC AAG AAC ACT GGC AGA GAG GAA TTC CTC ACA GCC TTC CTG CAG 146 Asp Leu Lys Asn Thr Gly Arg Glu Glu Phe Leu Thr Ala Phe Leu Gln 30 35 40 AAC TAT CAG CTG GCC TAC AGC AAG GCC TAC CCC CGC CTC CTT ATC TCC 194 Asn Tyr Gln Leu Ala Tyr Ser Lys Ala Tyr Pro Arg Leu Leu Ile Ser 45 50 55 AGT CTG TCA GAG AGC CCC GCT TCA GTC TCC ATC CTC AGC CAG GCA GAC 242 Ser Leu Ser Glu Ser Pro Ala Ser Val Ser Ile Leu Ser Gln Ala Asp 60 65 70 AAC ACC TCA AAG AAG GTC ACA GTG AGG CCC GGG GAG TCG GTC ATG GTC 290 Asn Thr Ser Lys Lys Val Thr Val Arg Pro Gly Glu Ser Val Met Val 75 80 85 90 AAC ATC AGT GCC AAG GCT GAG ATG ATA GGC AGC AAG ATC TTC CAG CAT 338 Asn Ile Ser Ala Lys Ala Glu Met Ile Gly Ser Lys Ile Phe Gln His 95 100 105 GCG GTG GTG ATC CAT TCT GAC TAT GCC ATC TCT GTG CAG GCA CTA AAT 386 Ala Val Val Ile His Ser Asp Tyr Ala Ile Ser Val Gln Ala Leu Asn 110 115 120 GCC AAG CCT GAC ACA GCG GAG CTG ACA CTG CTG CGG CCC ATC CAG GCC 434 Ala Lys Pro Asp Thr Ala Glu Leu Thr Leu Leu Arg Pro Ile Gln Ala 125 130 135 CTA GGC ACC GAG TAT TTT GTG CTC ACA CCC CCC GGC ACC TCA GCC AGG 482 Leu Gly Thr Glu Tyr Phe Val Leu Thr Pro Pro Gly Thr Ser Ala Arg 140 145 150 AAT GTC AAG GAG TTT GCC GTG GTG GCC GGT GCC GCA GGT GCC TCG GTC 530 Asn Val Lys Glu Phe Ala Val Val Ala Gly Ala Ala Gly Ala Ser Val 155 160 165 170 AGT GTC ACG CTG AAG GGG TCA GTG ACA TTC AAT GGC AAG TTC TAT CCA 578 Ser Val Thr Leu Lys Gly Ser Val Thr Phe Asn Gly Lys Phe Tyr Pro 175 180 185 GCA GGC GAT GTC CTA AGA GTG ACT CTA CAG CCC TAC AAT GTG GCC CAG 626 Ala Gly Asp Val Leu Arg Val Thr Leu Gln Pro Tyr Asn Val Ala Gln 190 195 200 CTA CAG AGC TCA GTG GAT CTC TCG GGG TCA AAG GTC ACA GCT AGT AGC 674 Leu Gln Ser Ser Val Asp Leu Ser Gly Ser Lys Val Thr Ala Ser Ser 205 210 215 CCC GTG GCT GTC CTC TCT GGC CAC AGC TGT GCG CAG AAA CAT ACG ACC 722 Pro Val Ala Val Leu Ser Gly His Ser Cys Ala Gln Lys His Thr Thr 220 225 230 TGC AAC CAT GTG GTT GAG CAG CTG CTA CCC ACG TCT GCC TGG GGC ACC 770 Cys Asn His Val Val Glu Gln Leu Leu Pro Thr Ser Ala Trp Gly Thr 235 240 245 250 CAC TAT GTA GTA CCC ACG CTG GCC TCC CAA TCT CGC TAT GAT TTG GCC 818 His Tyr Val Val Pro Thr Leu Ala Ser Gln Ser Arg Tyr Asp Leu Ala 255 260 265 TTC GTT GTG GCC AGC CAG GCC ACA AAG CTG ACC TAC AAC CAT GGG GGT 866 Phe Val Val Ala Ser Gln Ala Thr Lys Leu Thr Tyr Asn His Gly Gly 270 275 280 ATC ACT GGC TCC CGT GGG CTC CAG GCA GGT GAT GTG GTA GAG TTT GAG 914 Ile Thr Gly Ser Arg Gly Leu Gln Ala Gly Asp Val Val Glu Phe Glu 285 290 295 GTC CGG CCA TCC TGG CCA CTC TAC CTG TCT GCA AAT GTG GGC ATC CAG 962 Val Arg Pro Ser Trp Pro Leu Tyr Leu Ser Ala Asn Val Gly Ile Gln 300 305 310 GTC CTG TTG TTT GGC ACA GGT GCC ATA AGG AAT GAA GTG ACT TAT GAC 1010 Val Leu Leu Phe Gly Thr Gly Ala Ile Arg Asn Glu Val Thr Tyr Asp 315 320 325 330 CCC TAC CTG GTC CTG ATC CCA GAT GTG GCG GCC TAC TGC CCA GCC TAT 1058 Pro Tyr Leu Val Leu Ile Pro Asp Val Ala Ala Tyr Cys Pro Ala Tyr 335 340 345 GTG GTC AAG AGT GTA CCA GGC TGT GAG GGC GTG GCC CTG GTA GTG GCA 1106 Val Val Lys Ser Val Pro Gly Cys Glu Gly Val Ala Leu Val Val Ala 350 355 360 CAG ACG AAG GCT ATC AGC GGG CTG ACC ATA GAT GGG CAT GCA GTG GGG 1154 Gln Thr Lys Ala Ile Ser Gly Leu Thr Ile Asp Gly His Ala Val Gly 365 370 375 GCC AAG CTC ACC TGG GAG GCT GTG CCA GGC AGT GAG TTC TCG TAT GCT 1202 Ala Lys Leu Thr Trp Glu Ala Val Pro Gly Ser Glu Phe Ser Tyr Ala 380 385 390 GAA GTG GAG CTC GGC ACA GCT GAC ATG ATC CAC ACG GCC GAG GCC ACC 1250 Glu Val Glu Leu Gly Thr Ala Asp Met Ile His Thr Ala Glu Ala Thr 395 400 405 410 ACC AAC TTG GGA CTG CTC ACC TTC GGG CTG GCC AAG GCT ATA GGC TAC 1298 Thr Asn Leu Gly Leu Leu Thr Phe Gly Leu Ala Lys Ala Ile Gly Tyr 415 420 425 GCA ACA GCT GCT GAT TGC GGC CGG ACT GTA CTG TCC CCA GTG GAG CCC 1346 Ala Thr Ala Ala Asp Cys Gly Arg Thr Val Leu Ser Pro Val Glu Pro 430 435 440 TCC TGC GAA GGC ATG CAG TGC GCA GCC GGG CAG CGC TGC CAG GTG GTA 1394 Ser Cys Glu Gly Met Gln Cys Ala Ala Gly Gln Arg Cys Gln Val Val 445 450 455 GGC GGG AAG GCC GGG TGT GTG GCG GAG TCC ACC GCT GTC TGC CGC GCC 1442 Gly Gly Lys Ala Gly Cys Val Ala Glu Ser Thr Ala Val Cys Arg Ala 460 465 470 CAG GGC GAC CCC CAT TAC ACC ACC TTC GAC GGC CGT CGC TAC GAC ATG 1490 Gln Gly Asp Pro His Tyr Thr Thr Phe Asp Gly Arg Arg Tyr Asp Met 475 480 485 490 ATG GGC ACC TGT TCG TAC ACG ATG GTG GAG CTG TGC AGC GAG GAC GAC 1538 Met Gly Thr Cys Ser Tyr Thr Met Val Glu Leu Cys Ser Glu Asp Asp 495 500 505 ACC CTG CCC GCC TTC AGC GTG GAG GCC AAG AAC GAG CAC CGG GGC AGC 1586 Thr Leu Pro Ala Phe Ser Val Glu Ala Lys Asn Glu His Arg Gly Ser 510 515 520 CGC CGC GTC TCC TAC GTG GGC CTC GTC ACT GTG CGC GCC TAC AGC CAC 1634 Arg Arg Val Ser Tyr Val Gly Leu Val Thr Val Arg Ala Tyr Ser His 525 530 535 TCT GTG TCG CTG ACC CGC GGT GAA GTT GGC TTC GTC CTG GTT GAC AAC 1682 Ser Val Ser Leu Thr Arg Gly Glu Val Gly Phe Val Leu Val Asp Asn 540 545 550 CAG CGC TCG CGC CTG CCA GTC TCC CTG AGT GAG GGT CGC CTG CGT GTG 1730 Gln Arg Ser Arg Leu Pro Val Ser Leu Ser Glu Gly Arg Leu Arg Val 555 560 565 570 TAC CAG AGC GGA CCA CGG GCC GTG GTG GAG CTG GTC TTT GGG CTG GTG 1778 Tyr Gln Ser Gly Pro Arg Ala Val Val Glu Leu Val Phe Gly Leu Val 575 580 585 GTC ACT TAT GAC TGG GAC TGC CAG CTG GCA CTC AGC CTG CCT GCA CGC 1826 Val Thr Tyr Asp Trp Asp Cys Gln Leu Ala Leu Ser Leu Pro Ala Arg 590 595 600 TTC CAA GAC CAG GTG TGC GGG CTG TGT GGC AAC TAT AAT GGT GAC CCA 1874 Phe Gln Asp Gln Val Cys Gly Leu Cys Gly Asn Tyr Asn Gly Asp Pro 605 610 615 GCA GAC GAC TTC CTC ACG CCT GAC GGG GCT CTG GCT CCT GAC GCT GTG 1922 Ala Asp Asp Phe Leu Thr Pro Asp Gly Ala Leu Ala Pro Asp Ala Val 620 625 630 GAG TTC GCA AGT AGC TGG AAG CTG GAT GAT GGG GAC TAC CTG TGT GAG 1970 Glu Phe Ala Ser Ser Trp Lys Leu Asp Asp Gly Asp Tyr Leu Cys Glu 635 640 645 650 GAT GGC TGC CAG AAC AAC TGT CCC GCC TGC ACC CCA GGC CAG GCC CAA 2018 Asp Gly Cys Gln Asn Asn Cys Pro Ala Cys Thr Pro Gly Gln Ala Gln 655 660 665 CAC TAT GAG GGC GAC CGA CTC TGT GGC ATG CTG ACC AAG CTC GAT GGC 2066 His Tyr Glu Gly Asp Arg Leu Cys Gly Met Leu Thr Lys Leu Asp Gly 670 675 680 CCC TTC GCT GTC TGC CAT GAC ACC CTG GAC CCC AGG CCC TTC CTG GAG 2114 Pro Phe Ala Val Cys His Asp Thr Leu Asp Pro Arg Pro Phe Leu Glu 685 690 695 CAG TGT GTA TAT GAC CTG TGT GTG GTC GGT GGG GAG CGG CTC AGC CTG 2162 Gln Cys Val Tyr Asp Leu Cys Val Val Gly Gly Glu Arg Leu Ser Leu 700 705 710 TGC CGT GGC CTC AGC GCC TAT GCC CAG GCC TGT CTG GAG CTT GGC ATC 2210 Cys Arg Gly Leu Ser Ala Tyr Ala Gln Ala Cys Leu Glu Leu Gly Ile 715 720 725 730 TCG GTT GGG GAC TGG AGA TCA CCA GCC AAC TGC CCC CTG TCC TGC CCT 2258 Ser Val Gly Asp Trp Arg Ser Pro Ala Asn Cys Pro Leu Ser Cys Pro 735 740 745 GCC AAC AGC CGC TAT GAG CTC TGC GGC CCT GCT TGC CCG ACC TCC TGC 2306 Ala Asn Ser Arg Tyr Glu Leu Cys Gly Pro Ala Cys Pro Thr Ser Cys 750 755 760 AAC GGG GCT GCG GCG CCG TCC AAC TGC TCC GGG CGC CCC TGC GTG GAG 2354 Asn Gly Ala Ala Ala Pro Ser Asn Cys Ser Gly Arg Pro Cys Val Glu 765 770 775 GGC TGC GTG TGC CTC CCA GGC TTC GTG GCC AGC GGC GGC GCC TGC GTG 2402 Gly Cys Val Cys Leu Pro Gly Phe Val Ala Ser Gly Gly Ala Cys Val 780 785 790 CCG GCC TCG TCG TGT GGC TGC ACC TTC CAG GGT CTC CAG CTC GCT CCG 2450 Pro Ala Ser Ser Cys Gly Cys Thr Phe Gln Gly Leu Gln Leu Ala Pro 795 800 805 810 GGC CAG GAA GTG TGG GCG GAC GAG TTG TGC CAA AGG CGC TGC ACC TGC 2498 Gly Gln Glu Val Trp Ala Asp Glu Leu Cys Gln Arg Arg Cys Thr Cys 815 820 825 AAC GGC GCC ACC CAT CAG GTC ACC TGC CGC GAC AAG CAG AGC TGC CCG 2546 Asn Gly Ala Thr His Gln Val Thr Cys Arg Asp Lys Gln Ser Cys Pro 830 835 840 GCG GGT GAG CGC TGC AGC GTC CAG AAC GGC CTC CTG GGC TGC TAC CCC 2594 Ala Gly Glu Arg Cys Ser Val Gln Asn Gly Leu Leu Gly Cys Tyr Pro 845 850 855 GAT CGC TTC GGG ACC TGC CAG GGG TCC GGG GAC CCA CAC TAT GTG AGC 2642 Asp Arg Phe Gly Thr Cys Gln Gly Ser Gly Asp Pro His Tyr Val Ser 860 865 870 TTC GAC GGC CGG CGC TTC GAC TTC ATG GGC ACC TGC ACG TAC CTG CTG 2690 Phe Asp Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Thr Tyr Leu Leu 875 880 885 890 GTC GGC TCA TGC GGC CAG AAC GCA GCG CTG CCT GCC TTC CGG GTG CTG 2738 Val Gly Ser Cys Gly Gln Asn Ala Ala Leu Pro Ala Phe Arg Val Leu 895 900 905 GTG GAA AAC GAG CAT CGG GGC AGC CAG ACT GTG AGC TAC ACG CGC GCC 2786 Val Glu Asn Glu His Arg Gly Ser Gln Thr Val Ser Tyr Thr Arg Ala 910 915 920 GTG CGG GTG GAG GCC CGC GGG GTG AAG GTG GCC GTG CGC CGG GAG TAC 2834 Val Arg Val Glu Ala Arg Gly Val Lys Val Ala Val Arg Arg Glu Tyr 925 930 935 CCC GGG CAA GTG CTG GTG GAT GAC GTC CTT CAG TAT CTG CCC TTC CAA 2882 Pro Gly Gln Val Leu Val Asp Asp Val Leu Gln Tyr Leu Pro Phe Gln 940 945 950 GCA GCA GAT GGG CAG GTG CAG GTG TTC CGA CAG GGC AGG GAT GCC GTC 2930 Ala Ala Asp Gly Gln Val Gln Val Phe Arg Gln Gly Arg Asp Ala Val 955 960 965 970 GTG CGC ACG GAC TTT GGC CTG ACT GTC ACT TAT GAC TGG AAT GCA CGA 2978 Val Arg Thr Asp Phe Gly Leu Thr Val Thr Tyr Asp Trp Asn Ala Arg 975 980 985 GTG ACT GCC AAG GTG CCC AGC AGC TAT GCT GAG GCC CTG TGT GGA CTC 3026 Val Thr Ala Lys Val Pro Ser Ser Tyr Ala Glu Ala Leu Cys Gly Leu 990 995 1000 TGT GGG AAC TTC AAC GGG GAC CCA GCT GAT GAC CTG GCT CTG CGG GGT 3074 Cys Gly Asn Phe Asn Gly Asp Pro Ala Asp Asp Leu Ala Leu Arg Gly 1005 1010 1015 GGG GGT CAA GCT GCC AAT GCA CTG GCC TTT GGG AAC AGC TGG CAA GAA 3122 Gly Gly Gln Ala Ala Asn Ala Leu Ala Phe Gly Asn Ser Trp Gln Glu 1020 1025 1030 GAG ACG AGG CCC GGC TGT GGA GCA ACT GAA CCG GGT GAC TGT CCC AAG 3170 Glu Thr Arg Pro Gly Cys Gly Ala Thr Glu Pro Gly Asp Cys Pro Lys 1035 1040 1045 1050 CTG GAC TCC CTG GTG GCC CAG CAG CTG CAG AGC AAG AAT GAG TGT GGA 3218 Leu Asp Ser Leu Val Ala Gln Gln Leu Gln Ser Lys Asn Glu Cys Gly 1055 1060 1065 ATC CTT GCC GAC CCC AAG GGG CCC TTC CGG GAG TGC CAT AGC AAG CTG 3266 Ile Leu Ala Asp Pro Lys Gly Pro Phe Arg Glu Cys His Ser Lys Leu 1070 1075 1080 GAC CCC CAG GGT GCC GTG CGC GAC TGT GTC TAT GAC CGC TGC CTG CTG 3314 Asp Pro Gln Gly Ala Val Arg Asp Cys Val Tyr Asp Arg Cys Leu Leu 1085 1090 1095 CCA GGC CAG TCT GGG CCA CTG TGT GAC GCA CTG GCC ACC TAT GCT GCT 3362 Pro Gly Gln Ser Gly Pro Leu Cys Asp Ala Leu Ala Thr Tyr Ala Ala 1100 1105 1110 GCA TGC CAG GCT GCT GGA GCC ACA GTG CAC CCC TGG AGG AGT GAA GAA 3410 Ala Cys Gln Ala Ala Gly Ala Thr Val His Pro Trp Arg Ser Glu Glu 1115 1120 1125 1130 CTT TGC CCA CTG AGC TGC CCA CCC CAC AGC CAC TAT GAG GCG TGT TCC 3458 Leu Cys Pro Leu Ser Cys Pro Pro His Ser His Tyr Glu Ala Cys Ser 1135 1140 1145 TAC GGC TGC CCG CTG TCC TGT GGA GAC CTC CCA GTG CCC GGG GGC TGT 3506 Tyr Gly Cys Pro Leu Ser Cys Gly Asp Leu Pro Val Pro Gly Gly Cys 1150 1155 1160 GGC TCA GAA TGC CAT GAG GGC TGC GTG TGC GAT GAG GGC TTT GCG CTC 3554 Gly Ser Glu Cys His Glu Gly Cys Val Cys Asp Glu Gly Phe Ala Leu 1165 1170 1175 AGT GGT GAG TCC TGC CTG CCC CTG GCC TCC TGT GGC TGC GTA CAC CAG 3602 Ser Gly Glu Ser Cys Leu Pro Leu Ala Ser Cys Gly Cys Val His Gln 1180 1185 1190 GGC ACC TAC CAC CCA CCA GGC CAG ACC TTC TAC CCT GGC CCC GGA TGT 3650 Gly Thr Tyr His Pro Pro Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys 1195 1200 1205 1210 GAT TCC CTT TGC CAC TGC CAG GAG GGC GGC CTG GTG TCC TGT GAG TCC 3698 Asp Ser Leu Cys His Cys Gln Glu Gly Gly Leu Val Ser Cys Glu Ser 1215 1220 1225 TCC AGC TGC GGA CCG CAC GAG GCC TGC CAG CCA TCC GGT GGC AGC TTG 3746 Ser Ser Cys Gly Pro His Glu Ala Cys Gln Pro Ser Gly Gly Ser Leu 1230 1235 1240 GGC TGT GTG GCC GTG GGC TCT AGC ACC TGC CAG GCG TCA GGA GAC CCC 3794 Gly Cys Val Ala Val Gly Ser Ser Thr Cys Gln Ala Ser Gly Asp Pro 1245 1250 1255 CAC TAC ACC ACC TTC GAT GGC CGC CGC TTC GAC TTC ATG GGC ACC TGC 3842 His Tyr Thr Thr Phe Asp Gly Arg Arg Phe Asp Phe Met Gly Thr Cys 1260 1265 1270 GTG TAT GTG CTG GCT CAG ACC TGC GGC ACC CGG CCT GGC CTG CAT CGG 3890 Val Tyr Val Leu Ala Gln Thr Cys Gly Thr Arg Pro Gly Leu His Arg 1275 1280 1285 1290 TTT GCC GTC CTG CAG GAG AAC GTG GCC TGG GGT AAT GGG CGA GTC AGT 3938 Phe Ala Val Leu Gln Glu Asn Val Ala Trp Gly Asn Gly Arg Val Ser 1295 1300 1305 GTG ACC AGG GTG ATC ACG GTC CAG GTG GCA AAC TTC ACC CTG CGG CTG 3986 Val Thr Arg Val Ile Thr Val Gln Val Ala Asn Phe Thr Leu Arg Leu 1310 1315 1320 GAG CAG AGA CAG TGG AAG GTC ACG GTG AAC GGT GTG GAC ATG AAG CTG 4034 Glu Gln Arg Gln Trp Lys Val Thr Val Asn Gly Val Asp Met Lys Leu 1325 1330 1335 CCC GTG GTG CTG GCC AAC GGC CAG ATC CGT GCC TCC CAG CAT GGT TCA 4082 Pro Val Val Leu Ala Asn Gly Gln Ile Arg Ala Ser Gln His Gly Ser 1340 1345 1350 GAT GTT GTG ATT GAG ACC GAC TTC GGC CTG CGT GTG GCC TAC GAC CTT 4130 Asp Val Val Ile Glu Thr Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu 1355 1360 1365 1370 GTG TAC TAT GTG CGG GTC ACC GTC CCC GGA AAC TAC TAC CAG CAG ATG 4178 Val Tyr Tyr Val Arg Val Thr Val Pro Gly Asn Tyr Tyr Gln Gln Met 1375 1380 1385 TGT GGC CTG TGT GGG AAC TAC AAC GGC GAC CCC AAG GAT GAC TTC CAG 4226 Cys Gly Leu Cys Gly Asn Tyr Asn Gly Asp Pro Lys Asp Asp Phe Gln 1390 1395 1400 AAG CCC AAT GGC TCA CAG GCA GGC AAC GCC AAT GAG TTC GGC AAC TCC 4274 Lys Pro Asn Gly Ser Gln Ala Gly Asn Ala Asn Glu Phe Gly Asn Ser 1405 1410 1415 TGG GAG GAG GTG GTG CCC GAC TCT CCC TGC CTG CCG CCC ACC CCT TGC 4322 Trp Glu Glu Val Val Pro Asp Ser Pro Cys Leu Pro Pro Thr Pro Cys 1420 1425 1430 CCG CCG GGG AGC GAG GAC TGT ATC CCC AGC CAC AAG TGT CCT CCC GAG 4370 Pro Pro Gly Ser Glu Asp Cys Ile Pro Ser His Lys Cys Pro Pro Glu 1435 1440 1445 1450 CTG GAG AAG AAG TAT CAG AAG GAG GAG TTC TGT GGG CTC CTC TCC AGC 4418 Leu Glu Lys Lys Tyr Gln Lys Glu Glu Phe Cys Gly Leu Leu Ser Ser 1455 1460 1465 CCC ACA GGG CCA CTG TCC TCC TGC CAC AAG CTG GTG GAT CCC CAG GGT 4466 Pro Thr Gly Pro Leu Ser Ser Cys His Lys Leu Val Asp Pro Gln Gly 1470 1475 1480 CCC TTG AAA GAT TGC ATC TTT GAT CTC TGC CTG GGT GGT GGG AAC CTG 4514 Pro Leu Lys Asp Cys Ile Phe Asp Leu Cys Leu Gly Gly Gly Asn Leu 1485 1490 1495 AGC ATT CTC TGC AGC AAC ATC CAT GCC TAC GTG AGT GCT TGC CAG GCG 4562 Ser Ile Leu Cys Ser Asn Ile His Ala Tyr Val Ser Ala Cys Gln Ala 1500 1505 1510 GCT GGA GGC CAC GTG GAG CCC TGG AGG ACT GAA ACT TTC TGT CCC ATG 4610 Ala Gly Gly His Val Glu Pro Trp Arg Thr Glu Thr Phe Cys Pro Met 1515 1520 1525 1530 GAG TGC CCT CCG AAC AGT CAC TAC GAG CTC TGT GCG GAC ACC TGC TCC 4658 Glu Cys Pro Pro Asn Ser His Tyr Glu Leu Cys Ala Asp Thr Cys Ser 1535 1540 1545 CTG GGC TGC TCA GCT CTC AGT GCC CCT CCA CAG TGC CAG GAT GGG TGT 4706 Leu Gly Cys Ser Ala Leu Ser Ala Pro Pro Gln Cys Gln Asp Gly Cys 1550 1555 1560 GCT GAG GGC TGC CAG TGT GAC TCC GGC TTC CTC TAC AAT GGC CAA GCC 4754 Ala Glu Gly Cys Gln Cys Asp Ser Gly Phe Leu Tyr Asn Gly Gln Ala 1565 1570 1575 TGC GTG CCC ATC CAG CAA TGC GGC TGC TAC CAC AAT GGT GTC TAC TAT 4802 Cys Val Pro Ile Gln Gln Cys Gly Cys Tyr His Asn Gly Val Tyr Tyr 1580 1585 1590 GAG CCG GAG CAG ACA GTC CTC ATT GAC AAC TGT CGG CAG CAG TGC ACG 4850 Glu Pro Glu Gln Thr Val Leu Ile Asp Asn Cys Arg Gln Gln Cys Thr 1595 1600 1605 1610 TGC CAT GCG GGT AAA GGC ATG GTG TGC CAG GAA CAC AGC TGC AAG CCG 4898 Cys His Ala Gly Lys Gly Met Val Cys Gln Glu His Ser Cys Lys Pro 1615 1620 1625 GGG CAG GTG TGC CAG CCC TCC GGA GGC ATC CTG AGC TGC GTC ACC AAA 4946 Gly Gln Val Cys Gln Pro Ser Gly Gly Ile Leu Ser Cys Val Thr Lys 1630 1635 1640 GAC CCG TGC CAC GGC GTG ACA TGC CGG CCA CAG GAG ACA TGC AAG GAG 4994 Asp Pro Cys His Gly Val Thr Cys Arg Pro Gln Glu Thr Cys Lys Glu 1645 1650 1655 CAG GGT GGC CAG GGC GTG TGC CTG CCC AAC TAT GAG GCC ACG TGC TGG 5042 Gln Gly Gly Gln Gly Val Cys Leu Pro Asn Tyr Glu Ala Thr Cys Trp 1660 1665 1670 CTG TGG GGC GAC CCA CAC TAC CAC TCC TTC GAT GGC CGG AAG TTT GAC 5090 Leu Trp Gly Asp Pro His Tyr His Ser Phe Asp Gly Arg Lys Phe Asp 1675 1680 1685 1690 TTC CAG GGC ACC TGT AAC TAT GTG CTG GCA ACA ACT GGC TGC CCG GGG 5138 Phe Gln Gly Thr Cys Asn Tyr Val Leu Ala Thr Thr Gly Cys Pro Gly 1695 1700 1705 GTC AGC ACC CAG GGC CTG ACA CCC TTC ACC GTC ACC ACC AAG AAC CAG 5186 Val Ser Thr Gln Gly Leu Thr Pro Phe Thr Val Thr Thr Lys Asn Gln 1710 1715 1720 AAC CGG GGC AAC CCT GCT GTG TCC TAC GTG AGA GTC GTC ACC GTG GCT 5234 Asn Arg Gly Asn Pro Ala Val Ser Tyr Val Arg Val Val Thr Val Ala 1725 1730 1735 GCC CTC GGC ACC AAC ATC TCC ATC CAC AAG GAC GAG ATC GGC AAA GTC 5282 Ala Leu Gly Thr Asn Ile Ser Ile His Lys Asp Glu Ile Gly Lys Val 1740 1745 1750 CGG GTG AAC GGT GTG CTC ACA GCC TTG CCT GTC TCT GTG GCC GAC GGG 5330 Arg Val Asn Gly Val Leu Thr Ala Leu Pro Val Ser Val Ala Asp Gly 1755 1760 1765 1770 CGG ATT TCA GTG ACC CAG GGT GCA TCG AAG GCA CTG CTG GTG GCT GAC 5378 Arg Ile Ser Val Thr Gln Gly Ala Ser Lys Ala Leu Leu Val Ala Asp 1775 1780 1785 TTT GGA CTG CAA GTC AGC TAT GAC TGG AAC TGG CGG GTA GAC GTG ACG 5426 Phe Gly Leu Gln Val Ser Tyr Asp Trp Asn Trp Arg Val Asp Val Thr 1790 1795 1800 CTG CCC AGC AGC TAT CAT GGC GCA GTG TGC GGG CTC TGC GGT AAC ATG 5474 Leu Pro Ser Ser Tyr His Gly Ala Val Cys Gly Leu Cys Gly Asn Met 1805 1810 1815 GAC CGC AAC CCC AAC AAT GAC CAG GTC TTC CCT AAT GGC ACA CTG GCT 5522 Asp Arg Asn Pro Asn Asn Asp Gln Val Phe Pro Asn Gly Thr Leu Ala 1820 1825 1830 CCC TCC ATA CCC ATC TGG GGC GGC AGC TGG CGA GCC CCA GGC TGG GAC 5570 Pro Ser Ile Pro Ile Trp Gly Gly Ser Trp Arg Ala Pro Gly Trp Asp 1835 1840 1845 1850 CCA CTG TGT TGG GAC GAA TGT CGG GGG TCC TGC CCA ACG TGC CCT GAG 5618 Pro Leu Cys Trp Asp Glu Cys Arg Gly Ser Cys Pro Thr Cys Pro Glu 1855 1860 1865 GAC CGG TTG GAG CAG TAC GAG GGC CCT GGC TTC TGC GGA CCC CTG GCC 5666 Asp Arg Leu Glu Gln Tyr Glu Gly Pro Gly Phe Cys Gly Pro Leu Ala 1870 1875 1880 CCC GGC ACA GGG GGC CCT TTC ACC ACC TGC CAT GCT CAT GTG CCA CCT 5714 Pro Gly Thr Gly Gly Pro Phe Thr Thr Cys His Ala His Val Pro Pro 1885 1890 1895 GAG AGC TTC TTC AAG GGC TGT GTT CTG GAC GTC TGC ATG GGT GGT GGG 5762 Glu Ser Phe Phe Lys Gly Cys Val Leu Asp Val Cys Met Gly Gly Gly 1900 1905 1910 GAC CGT GAC ATT CTT TGC AAG GCT CTG GCT TCC TAT GTG GCC GCC TGC 5810 Asp Arg Asp Ile Leu Cys Lys Ala Leu Ala Ser Tyr Val Ala Ala Cys 1915 1920 1925 1930 CAG GCT GCT GGG GTT GTC ATC GAA GAC TGG CGG GCA CAG GTT GGC TGT 5858 Gln Ala Ala Gly Val Val Ile Glu Asp Trp Arg Ala Gln Val Gly Cys 1935 1940 1945 GAG ATC ACC TGC CCA GAA AAC AGC CAC TAT GAG GTC TGT GGC CCA CCC 5906 Glu Ile Thr Cys Pro Glu Asn Ser His Tyr Glu Val Cys Gly Pro Pro 1950 1955 1960 TGC CCG GCC AGC TGT CCG TCC CCT GCA CCC CTT ACG ACG CCA GCC GTA 5954 Cys Pro Ala Ser Cys Pro Ser Pro Ala Pro Leu Thr Thr Pro Ala Val 1965 1970 1975 TGT GAG GGC CCC TGT GTG GAG GGC TGC CAG TGC GAC GCG GGT TTC GTG 6002 Cys Glu Gly Pro Cys Val Glu Gly Cys Gln Cys Asp Ala Gly Phe Val 1980 1985 1990 TTA AGT GCT GAC CGC TGT GTT CCC CTC AAC AAC GGC TGC GGC TGC TGG 6050 Leu Ser Ala Asp Arg Cys Val Pro Leu Asn Asn Gly Cys Gly Cys Trp 1995 2000 2005 2010 GCC AAT GGC ACC TAC CAC GAG GCG GGC AGT GAG TTT TGG GCT GAT GGC 6098 Ala Asn Gly Thr Tyr His Glu Ala Gly Ser Glu Phe Trp Ala Asp Gly 2015 2020 2025 ACC TGC TCC CAG TGG TGT CGC TGC GGG CCT GGG GGT GGC TCG CTG GTC 6146 Thr Cys Ser Gln Trp Cys Arg Cys Gly Pro Gly Gly Gly Ser Leu Val 2030 2035 2040 TGC ACA CCT GCC AGC TGT GGG CTG GGT GAA GTG TGT GGC CTC CTG CCA 6194 Cys Thr Pro Ala Ser Cys Gly Leu Gly Glu Val Cys Gly Leu Leu Pro 2045 2050 2055 TCC GGC CAG CAC GGC TGC CAG CCC GTC AGC ACA GCT GAG TGC CAG GCG 6242 Ser Gly Gln His Gly Cys Gln Pro Val Ser Thr Ala Glu Cys Gln Ala 2060 2065 2070 TGG GGT GAC CCC CAT TAC GTC ACT CTG GAT GGG CAC CGA TTC AAT TTC 6290 Trp Gly Asp Pro His Tyr Val Thr Leu Asp Gly His Arg Phe Asn Phe 2075 2080 2085 2090 CAA GGC ACC TGC GAG TAC CTG CTG AGT GCA CCC TGC CAC GGA CCA CCC 6338 Gln Gly Thr Cys Glu Tyr Leu Leu Ser Ala Pro Cys His Gly Pro Pro 2095 2100 2105 TTG GGG GCT GAG AAC TTC ACT GTC ACT GTA GCC AAT GAG CAC CGG GGC 6386 Leu Gly Ala Glu Asn Phe Thr Val Thr Val Ala Asn Glu His Arg Gly 2110 2115 2120 AGC CAG GCT GTC AGC TAC ACC CGC AGT GTC ACC CTG CAA ATC TAC AAC 6434 Ser Gln Ala Val Ser Tyr Thr Arg Ser Val Thr Leu Gln Ile Tyr Asn 2125 2130 2135 CAC AGC CTG ACA CTG AGT GCC CGC TGG CCC CGG AAG CTA CAG GTG GAC 6482 His Ser Leu Thr Leu Ser Ala Arg Trp Pro Arg Lys Leu Gln Val Asp 2140 2145 2150 GGC GTG TTC GTC ACT CTG CCC TTC CAG CTG GAC TCG CTC CTG CAC GCA 6530 Gly Val Phe Val Thr Leu Pro Phe Gln Leu Asp Ser Leu Leu His Ala 2155 2160 2165 2170 CAC CTG AGC GGC GCC GAC GTG GTG GTG ACC ACA ACC TCA GGG CTC TCG 6578 His Leu Ser Gly Ala Asp Val Val Val Thr Thr Thr Ser Gly Leu Ser 2175 2180 2185 CTG GCT TTC GAC GGG GAC AGC TTC GTG CGC CTG CGC GTG CCG GCG GCG 6626 Leu Ala Phe Asp Gly Asp Ser Phe Val Arg Leu Arg Val Pro Ala Ala 2190 2195 2200 TAC GCG GGC TCT CTC TGT GGC TTA TGC GGG AAC TAC AAC CAG GAC CCC 6674 Tyr Ala Gly Ser Leu Cys Gly Leu Cys Gly Asn Tyr Asn Gln Asp Pro 2205 2210 2215 GCA GAC GAC CTG AAG GCG GTG GGC GGG AAG CCC GCC GGA TGG CAG GTG 6722 Ala Asp Asp Leu Lys Ala Val Gly Gly Lys Pro Ala Gly Trp Gln Val 2220 2225 2230 GGC GGC GCC CAG GGC TGC GGG GAA TGT GTG TCC AAG CCA TGC CCG TCG 6770 Gly Gly Ala Gln Gly Cys Gly Glu Cys Val Ser Lys Pro Cys Pro Ser 2235 2240 2245 2250 CCG TGC ACC CCA GAG CAG CAA GAG TCC TTC GGC GGC CCG GAC GCC TGC 6818 Pro Cys Thr Pro Glu Gln Gln Glu Ser Phe Gly Gly Pro Asp Ala Cys 2255 2260 2265 GGC GTG ATC TCC GCC ACC GAC GGC CCG CTG GCG CCC TGC CAC GGC CTT 6866 Gly Val Ile Ser Ala Thr Asp Gly Pro Leu Ala Pro Cys His Gly Leu 2270 2275 2280 GTG CCG CCC GCG CAG TAC TTC CAG GGC TGC TTG CTG GAC GCC TGC CAA 6914 Val Pro Pro Ala Gln Tyr Phe Gln Gly Cys Leu Leu Asp Ala Cys Gln 2285 2290 2295 GTT CAG GGC CAT CCT GGA GGC CTC TGT CCT GCA GTG GCC ACC TAC GTG 6962 Val Gln Gly His Pro Gly Gly Leu Cys Pro Ala Val Ala Thr Tyr Val 2300 2305 2310 GCA GCC TGT CAG GCC GCT GGG GCC CAG CTC CGC GAG TGG AGG CGG CCG 7010 Ala Ala Cys Gln Ala Ala Gly Ala Gln Leu Arg Glu Trp Arg Arg Pro 2315 2320 2325 2330 GAC TTC TGT CCC TTC CAG TGC CCT GCC CAC AGC CAC TAC GAG CTC TGC 7058 Asp Phe Cys Pro Phe Gln Cys Pro Ala His Ser His Tyr Glu Leu Cys 2335 2340 2345 GGT GAC TCC TGT CCT GGG AGC TGC CCG AGC CTG TCG GCA CCC GAG GGC 7106 Gly Asp Ser Cys Pro Gly Ser Cys Pro Ser Leu Ser Ala Pro Glu Gly 2350 2355 2360 TGT GAG TCG GCC TGC CGT GAA GGC TGT GTC TGC GAT GCT GGC TTC GTG 7154 Cys Glu Ser Ala Cys Arg Glu Gly Cys Val Cys Asp Ala Gly Phe Val 2365 2370 2375 CTC AGT GGT GAC ACG TGT GTA CCT GTG GGC CAG TGT GGC TGC CTC CAC 7202 Leu Ser Gly Asp Thr Cys Val Pro Val Gly Gln Cys Gly Cys Leu His 2380 2385 2390 GAT GAC CGC TAC TAC CCA CTG GGC CAG ACC TTC TAC CCT GGC CCT GGG 7250 Asp Asp Arg Tyr Tyr Pro Leu Gly Gln Thr Phe Tyr Pro Gly Pro Gly 2395 2400 2405 2410 TGT GAT TCC CTT TGC CGC TGC CGG GAG GGC GGT GAG GTG TCC TGT GAG 7298 Cys Asp Ser Leu Cys Arg Cys Arg Glu Gly Gly Glu Val Ser Cys Glu 2415 2420 2425 CCC TCC AGC TGC GGC CCG CAT GAG ACC TGC CGG CCA TCC GGT GGC AGC 7346 Pro Ser Ser Cys Gly Pro His Glu Thr Cys Arg Pro Ser Gly Gly Ser 2430 2435 2440 TTG GGC TGC GTG GCC GTG GGC TCT ACC ACC TGC CAG GCG TCG GGA GAT 7394 Leu Gly Cys Val Ala Val Gly Ser Thr Thr Cys Gln Ala Ser Gly Asp 2445 2450 2455 CCC CAC TAC ACC ACC TTC GAT GGC CGC CGC TTC GAC TTC ATG GGC ACC 7442 Pro His Tyr Thr Thr Phe Asp Gly Arg Arg Phe Asp Phe Met Gly Thr 2460 2465 2470 TGC GTG TAT GTG CTG GCT CAG ACC TGC GGC ACC CGG CCT GGC CTA CAT 7490 Cys Val Tyr Val Leu Ala Gln Thr Cys Gly Thr Arg Pro Gly Leu His 2475 2480 2485 2490 CGG TTT GCC GTC CTG CAG GAG AAC GTG GCC TGG GGT AAT GGG CGA GTC 7538 Arg Phe Ala Val Leu Gln Glu Asn Val Ala Trp Gly Asn Gly Arg Val 2495 2500 2505 AGT GTG ACC AGG GTG ATC ACG GTC CAG GTG GCA AAC TTC ACC CTG CGG 7586 Ser Val Thr Arg Val Ile Thr Val Gln Val Ala Asn Phe Thr Leu Arg 2510 2515 2520 CTG GAG CAG AGA CAG TGG AAG GTC ACG GTG AAC GGT GTG GAC ATG AAG 7634 Leu Glu Gln Arg Gln Trp Lys Val Thr Val Asn Gly Val Asp Met Lys 2525 2530 2535 CTG CCC GTG GTG CTG GCC AAC GGC CAG ATC CGT GCC TCC CAG CAT GGT 7682 Leu Pro Val Val Leu Ala Asn Gly Gln Ile Arg Ala Ser Gln His Gly 2540 2545 2550 TCA GAT GTT GTG ATT GAG ACC GAC TTC GGC CTG CGT GTG GCC TAC GAC 7730 Ser Asp Val Val Ile Glu Thr Asp Phe Gly Leu Arg Val Ala Tyr Asp 2555 2560 2565 2570 CTT GTG TAC TAT GTG CGG GTC ACC GTC CCT GGA AAC TAC TAC CAG CTG 7778 Leu Val Tyr Tyr Val Arg Val Thr Val Pro Gly Asn Tyr Tyr Gln Leu 2575 2580 2585 ATG TGT GGC CTG TGT GGG GGA TCC ACTAGTTAGT TAGTTAGGGT AC 7824 Met Cys Gly Leu Cys Gly Gly Ser 2590 2594 amino acids amino acid linear protein not provided 7 Met Gly Ala Leu Trp Ser Trp Trp Ile Leu Trp Ala Gly Ala Thr Leu 1 5 10 15 Leu Trp Gly Leu Thr Gln Glu Ala Ser Val Asp Leu Lys Asn Thr Gly 20 25 30 Arg Glu Glu Phe Leu Thr Ala Phe Leu Gln Asn Tyr Gln Leu Ala Tyr 35 40 45 Ser Lys Ala Tyr Pro Arg Leu Leu Ile Ser Ser Leu Ser Glu Ser Pro 50 55 60 Ala Ser Val Ser Ile Leu Ser Gln Ala Asp Asn Thr Ser Lys Lys Val 65 70 75 80 Thr Val Arg Pro Gly Glu Ser Val Met Val Asn Ile Ser Ala Lys Ala 85 90 95 Glu Met Ile Gly Ser Lys Ile Phe Gln His Ala Val Val Ile His Ser 100 105 110 Asp Tyr Ala Ile Ser Val Gln Ala Leu Asn Ala Lys Pro Asp Thr Ala 115 120 125 Glu Leu Thr Leu Leu Arg Pro Ile Gln Ala Leu Gly Thr Glu Tyr Phe 130 135 140 Val Leu Thr Pro Pro Gly Thr Ser Ala Arg Asn Val Lys Glu Phe Ala 145 150 155 160 Val Val Ala Gly Ala Ala Gly Ala Ser Val Ser Val Thr Leu Lys Gly 165 170 175 Ser Val Thr Phe Asn Gly Lys Phe Tyr Pro Ala Gly Asp Val Leu Arg 180 185 190 Val Thr Leu Gln Pro Tyr Asn Val Ala Gln Leu Gln Ser Ser Val Asp 195 200 205 Leu Ser Gly Ser Lys Val Thr Ala Ser Ser Pro Val Ala Val Leu Ser 210 215 220 Gly His Ser Cys Ala Gln Lys His Thr Thr Cys Asn His Val Val Glu 225 230 235 240 Gln Leu Leu Pro Thr Ser Ala Trp Gly Thr His Tyr Val Val Pro Thr 245 250 255 Leu Ala Ser Gln Ser Arg Tyr Asp Leu Ala Phe Val Val Ala Ser Gln 260 265 270 Ala Thr Lys Leu Thr Tyr Asn His Gly Gly Ile Thr Gly Ser Arg Gly 275 280 285 Leu Gln Ala Gly Asp Val Val Glu Phe Glu Val Arg Pro Ser Trp Pro 290 295 300 Leu Tyr Leu Ser Ala Asn Val Gly Ile Gln Val Leu Leu Phe Gly Thr 305 310 315 320 Gly Ala Ile Arg Asn Glu Val Thr Tyr Asp Pro Tyr Leu Val Leu Ile 325 330 335 Pro Asp Val Ala Ala Tyr Cys Pro Ala Tyr Val Val Lys Ser Val Pro 340 345 350 Gly Cys Glu Gly Val Ala Leu Val Val Ala Gln Thr Lys Ala Ile Ser 355 360 365 Gly Leu Thr Ile Asp Gly His Ala Val Gly Ala Lys Leu Thr Trp Glu 370 375 380 Ala Val Pro Gly Ser Glu Phe Ser Tyr Ala Glu Val Glu Leu Gly Thr 385 390 395 400 Ala Asp Met Ile His Thr Ala Glu Ala Thr Thr Asn Leu Gly Leu Leu 405 410 415 Thr Phe Gly Leu Ala Lys Ala Ile Gly Tyr Ala Thr Ala Ala Asp Cys 420 425 430 Gly Arg Thr Val Leu Ser Pro Val Glu Pro Ser Cys Glu Gly Met Gln 435 440 445 Cys Ala Ala Gly Gln Arg Cys Gln Val Val Gly Gly Lys Ala Gly Cys 450 455 460 Val Ala Glu Ser Thr Ala Val Cys Arg Ala Gln Gly Asp Pro His Tyr 465 470 475 480 Thr Thr Phe Asp Gly Arg Arg Tyr Asp Met Met Gly Thr Cys Ser Tyr 485 490 495 Thr Met Val Glu Leu Cys Ser Glu Asp Asp Thr Leu Pro Ala Phe Ser 500 505 510 Val Glu Ala Lys Asn Glu His Arg Gly Ser Arg Arg Val Ser Tyr Val 515 520 525 Gly Leu Val Thr Val Arg Ala Tyr Ser His Ser Val Ser Leu Thr Arg 530 535 540 Gly Glu Val Gly Phe Val Leu Val Asp Asn Gln Arg Ser Arg Leu Pro 545 550 555 560 Val Ser Leu Ser Glu Gly Arg Leu Arg Val Tyr Gln Ser Gly Pro Arg 565 570 575 Ala Val Val Glu Leu Val Phe Gly Leu Val Val Thr Tyr Asp Trp Asp 580 585 590 Cys Gln Leu Ala Leu Ser Leu Pro Ala Arg Phe Gln Asp Gln Val Cys 595 600 605 Gly Leu Cys Gly Asn Tyr Asn Gly Asp Pro Ala Asp Asp Phe Leu Thr 610 615 620 Pro Asp Gly Ala Leu Ala Pro Asp Ala Val Glu Phe Ala Ser Ser Trp 625 630 635 640 Lys Leu Asp Asp Gly Asp Tyr Leu Cys Glu Asp Gly Cys Gln Asn Asn 645 650 655 Cys Pro Ala Cys Thr Pro Gly Gln Ala Gln His Tyr Glu Gly Asp Arg 660 665 670 Leu Cys Gly Met Leu Thr Lys Leu Asp Gly Pro Phe Ala Val Cys His 675 680 685 Asp Thr Leu Asp Pro Arg Pro Phe Leu Glu Gln Cys Val Tyr Asp Leu 690 695 700 Cys Val Val Gly Gly Glu Arg Leu Ser Leu Cys Arg Gly Leu Ser Ala 705 710 715 720 Tyr Ala Gln Ala Cys Leu Glu Leu Gly Ile Ser Val Gly Asp Trp Arg 725 730 735 Ser Pro Ala Asn Cys Pro Leu Ser Cys Pro Ala Asn Ser Arg Tyr Glu 740 745 750 Leu Cys Gly Pro Ala Cys Pro Thr Ser Cys Asn Gly Ala Ala Ala Pro 755 760 765 Ser Asn Cys Ser Gly Arg Pro Cys Val Glu Gly Cys Val Cys Leu Pro 770 775 780 Gly Phe Val Ala Ser Gly Gly Ala Cys Val Pro Ala Ser Ser Cys Gly 785 790 795 800 Cys Thr Phe Gln Gly Leu Gln Leu Ala Pro Gly Gln Glu Val Trp Ala 805 810 815 Asp Glu Leu Cys Gln Arg Arg Cys Thr Cys Asn Gly Ala Thr His Gln 820 825 830 Val Thr Cys Arg Asp Lys Gln Ser Cys Pro Ala Gly Glu Arg Cys Ser 835 840 845 Val Gln Asn Gly Leu Leu Gly Cys Tyr Pro Asp Arg Phe Gly Thr Cys 850 855 860 Gln Gly Ser Gly Asp Pro His Tyr Val Ser Phe Asp Gly Arg Arg Phe 865 870 875 880 Asp Phe Met Gly Thr Cys Thr Tyr Leu Leu Val Gly Ser Cys Gly Gln 885 890 895 Asn Ala Ala Leu Pro Ala Phe Arg Val Leu Val Glu Asn Glu His Arg 900 905 910 Gly Ser Gln Thr Val Ser Tyr Thr Arg Ala Val Arg Val Glu Ala Arg 915 920 925 Gly Val Lys Val Ala Val Arg Arg Glu Tyr Pro Gly Gln Val Leu Val 930 935 940 Asp Asp Val Leu Gln Tyr Leu Pro Phe Gln Ala Ala Asp Gly Gln Val 945 950 955 960 Gln Val Phe Arg Gln Gly Arg Asp Ala Val Val Arg Thr Asp Phe Gly 965 970 975 Leu Thr Val Thr Tyr Asp Trp Asn Ala Arg Val Thr Ala Lys Val Pro 980 985 990 Ser Ser Tyr Ala Glu Ala Leu Cys Gly Leu Cys Gly Asn Phe Asn Gly 995 1000 1005 Asp Pro Ala Asp Asp Leu Ala Leu Arg Gly Gly Gly Gln Ala Ala Asn 1010 1015 1020 Ala Leu Ala Phe Gly Asn Ser Trp Gln Glu Glu Thr Arg Pro Gly Cys 1025 1030 1035 1040 Gly Ala Thr Glu Pro Gly Asp Cys Pro Lys Leu Asp Ser Leu Val Ala 1045 1050 1055 Gln Gln Leu Gln Ser Lys Asn Glu Cys Gly Ile Leu Ala Asp Pro Lys 1060 1065 1070 Gly Pro Phe Arg Glu Cys His Ser Lys Leu Asp Pro Gln Gly Ala Val 1075 1080 1085 Arg Asp Cys Val Tyr Asp Arg Cys Leu Leu Pro Gly Gln Ser Gly Pro 1090 1095 1100 Leu Cys Asp Ala Leu Ala Thr Tyr Ala Ala Ala Cys Gln Ala Ala Gly 1105 1110 1115 1120 Ala Thr Val His Pro Trp Arg Ser Glu Glu Leu Cys Pro Leu Ser Cys 1125 1130 1135 Pro Pro His Ser His Tyr Glu Ala Cys Ser Tyr Gly Cys Pro Leu Ser 1140 1145 1150 Cys Gly Asp Leu Pro Val Pro Gly Gly Cys Gly Ser Glu Cys His Glu 1155 1160 1165 Gly Cys Val Cys Asp Glu Gly Phe Ala Leu Ser Gly Glu Ser Cys Leu 1170 1175 1180 Pro Leu Ala Ser Cys Gly Cys Val His Gln Gly Thr Tyr His Pro Pro 1185 1190 1195 1200 Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys Asp Ser Leu Cys His Cys 1205 1210 1215 Gln Glu Gly Gly Leu Val Ser Cys Glu Ser Ser Ser Cys Gly Pro His 1220 1225 1230 Glu Ala Cys Gln Pro Ser Gly Gly Ser Leu Gly Cys Val Ala Val Gly 1235 1240 1245 Ser Ser Thr Cys Gln Ala Ser Gly Asp Pro His Tyr Thr Thr Phe Asp 1250 1255 1260 Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Val Tyr Val Leu Ala Gln 1265 1270 1275 1280 Thr Cys Gly Thr Arg Pro Gly Leu His Arg Phe Ala Val Leu Gln Glu 1285 1290 1295 Asn Val Ala Trp Gly Asn Gly Arg Val Ser Val Thr Arg Val Ile Thr 1300 1305 1310 Val Gln Val Ala Asn Phe Thr Leu Arg Leu Glu Gln Arg Gln Trp Lys 1315 1320 1325 Val Thr Val Asn Gly Val Asp Met Lys Leu Pro Val Val Leu Ala Asn 1330 1335 1340 Gly Gln Ile Arg Ala Ser Gln His Gly Ser Asp Val Val Ile Glu Thr 1345 1350 1355 1360 Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu Val Tyr Tyr Val Arg Val 1365 1370 1375 Thr Val Pro Gly Asn Tyr Tyr Gln Gln Met Cys Gly Leu Cys Gly Asn 1380 1385 1390 Tyr Asn Gly Asp Pro Lys Asp Asp Phe Gln Lys Pro Asn Gly Ser Gln 1395 1400 1405 Ala Gly Asn Ala Asn Glu Phe Gly Asn Ser Trp Glu Glu Val Val Pro 1410 1415 1420 Asp Ser Pro Cys Leu Pro Pro Thr Pro Cys Pro Pro Gly Ser Glu Asp 1425 1430 1435 1440 Cys Ile Pro Ser His Lys Cys Pro Pro Glu Leu Glu Lys Lys Tyr Gln 1445 1450 1455 Lys Glu Glu Phe Cys Gly Leu Leu Ser Ser Pro Thr Gly Pro Leu Ser 1460 1465 1470 Ser Cys His Lys Leu Val Asp Pro Gln Gly Pro Leu Lys Asp Cys Ile 1475 1480 1485 Phe Asp Leu Cys Leu Gly Gly Gly Asn Leu Ser Ile Leu Cys Ser Asn 1490 1495 1500 Ile His Ala Tyr Val Ser Ala Cys Gln Ala Ala Gly Gly His Val Glu 1505 1510 1515 1520 Pro Trp Arg Thr Glu Thr Phe Cys Pro Met Glu Cys Pro Pro Asn Ser 1525 1530 1535 His Tyr Glu Leu Cys Ala Asp Thr Cys Ser Leu Gly Cys Ser Ala Leu 1540 1545 1550 Ser Ala Pro Pro Gln Cys Gln Asp Gly Cys Ala Glu Gly Cys Gln Cys 1555 1560 1565 Asp Ser Gly Phe Leu Tyr Asn Gly Gln Ala Cys Val Pro Ile Gln Gln 1570 1575 1580 Cys Gly Cys Tyr His Asn Gly Val Tyr Tyr Glu Pro Glu Gln Thr Val 1585 1590 1595 1600 Leu Ile Asp Asn Cys Arg Gln Gln Cys Thr Cys His Ala Gly Lys Gly 1605 1610 1615 Met Val Cys Gln Glu His Ser Cys Lys Pro Gly Gln Val Cys Gln Pro 1620 1625 1630 Ser Gly Gly Ile Leu Ser Cys Val Thr Lys Asp Pro Cys His Gly Val 1635 1640 1645 Thr Cys Arg Pro Gln Glu Thr Cys Lys Glu Gln Gly Gly Gln Gly Val 1650 1655 1660 Cys Leu Pro Asn Tyr Glu Ala Thr Cys Trp Leu Trp Gly Asp Pro His 1665 1670 1675 1680 Tyr His Ser Phe Asp Gly Arg Lys Phe Asp Phe Gln Gly Thr Cys Asn 1685 1690 1695 Tyr Val Leu Ala Thr Thr Gly Cys Pro Gly Val Ser Thr Gln Gly Leu 1700 1705 1710 Thr Pro Phe Thr Val Thr Thr Lys Asn Gln Asn Arg Gly Asn Pro Ala 1715 1720 1725 Val Ser Tyr Val Arg Val Val Thr Val Ala Ala Leu Gly Thr Asn Ile 1730 1735 1740 Ser Ile His Lys Asp Glu Ile Gly Lys Val Arg Val Asn Gly Val Leu 1745 1750 1755 1760 Thr Ala Leu Pro Val Ser Val Ala Asp Gly Arg Ile Ser Val Thr Gln 1765 1770 1775 Gly Ala Ser Lys Ala Leu Leu Val Ala Asp Phe Gly Leu Gln Val Ser 1780 1785 1790 Tyr Asp Trp Asn Trp Arg Val Asp Val Thr Leu Pro Ser Ser Tyr His 1795 1800 1805 Gly Ala Val Cys Gly Leu Cys Gly Asn Met Asp Arg Asn Pro Asn Asn 1810 1815 1820 Asp Gln Val Phe Pro Asn Gly Thr Leu Ala Pro Ser Ile Pro Ile Trp 1825 1830 1835 1840 Gly Gly Ser Trp Arg Ala Pro Gly Trp Asp Pro Leu Cys Trp Asp Glu 1845 1850 1855 Cys Arg Gly Ser Cys Pro Thr Cys Pro Glu Asp Arg Leu Glu Gln Tyr 1860 1865 1870 Glu Gly Pro Gly Phe Cys Gly Pro Leu Ala Pro Gly Thr Gly Gly Pro 1875 1880 1885 Phe Thr Thr Cys His Ala His Val Pro Pro Glu Ser Phe Phe Lys Gly 1890 1895 1900 Cys Val Leu Asp Val Cys Met Gly Gly Gly Asp Arg Asp Ile Leu Cys 1905 1910 1915 1920 Lys Ala Leu Ala Ser Tyr Val Ala Ala Cys Gln Ala Ala Gly Val Val 1925 1930 1935 Ile Glu Asp Trp Arg Ala Gln Val Gly Cys Glu Ile Thr Cys Pro Glu 1940 1945 1950 Asn Ser His Tyr Glu Val Cys Gly Pro Pro Cys Pro Ala Ser Cys Pro 1955 1960 1965 Ser Pro Ala Pro Leu Thr Thr Pro Ala Val Cys Glu Gly Pro Cys Val 1970 1975 1980 Glu Gly Cys Gln Cys Asp Ala Gly Phe Val Leu Ser Ala Asp Arg Cys 1985 1990 1995 2000 Val Pro Leu Asn Asn Gly Cys Gly Cys Trp Ala Asn Gly Thr Tyr His 2005 2010 2015 Glu Ala Gly Ser Glu Phe Trp Ala Asp Gly Thr Cys Ser Gln Trp Cys 2020 2025 2030 Arg Cys Gly Pro Gly Gly Gly Ser Leu Val Cys Thr Pro Ala Ser Cys 2035 2040 2045 Gly Leu Gly Glu Val Cys Gly Leu Leu Pro Ser Gly Gln His Gly Cys 2050 2055 2060 Gln Pro Val Ser Thr Ala Glu Cys Gln Ala Trp Gly Asp Pro His Tyr 2065 2070 2075 2080 Val Thr Leu Asp Gly His Arg Phe Asn Phe Gln Gly Thr Cys Glu Tyr 2085 2090 2095 Leu Leu Ser Ala Pro Cys His Gly Pro Pro Leu Gly Ala Glu Asn Phe 2100 2105 2110 Thr Val Thr Val Ala Asn Glu His Arg Gly Ser Gln Ala Val Ser Tyr 2115 2120 2125 Thr Arg Ser Val Thr Leu Gln Ile Tyr Asn His Ser Leu Thr Leu Ser 2130 2135 2140 Ala Arg Trp Pro Arg Lys Leu Gln Val Asp Gly Val Phe Val Thr Leu 2145 2150 2155 2160 Pro Phe Gln Leu Asp Ser Leu Leu His Ala His Leu Ser Gly Ala Asp 2165 2170 2175 Val Val Val Thr Thr Thr Ser Gly Leu Ser Leu Ala Phe Asp Gly Asp 2180 2185 2190 Ser Phe Val Arg Leu Arg Val Pro Ala Ala Tyr Ala Gly Ser Leu Cys 2195 2200 2205 Gly Leu Cys Gly Asn Tyr Asn Gln Asp Pro Ala Asp Asp Leu Lys Ala 2210 2215 2220 Val Gly Gly Lys Pro Ala Gly Trp Gln Val Gly Gly Ala Gln Gly Cys 2225 2230 2235 2240 Gly Glu Cys Val Ser Lys Pro Cys Pro Ser Pro Cys Thr Pro Glu Gln 2245 2250 2255 Gln Glu Ser Phe Gly Gly Pro Asp Ala Cys Gly Val Ile Ser Ala Thr 2260 2265 2270 Asp Gly Pro Leu Ala Pro Cys His Gly Leu Val Pro Pro Ala Gln Tyr 2275 2280 2285 Phe Gln Gly Cys Leu Leu Asp Ala Cys Gln Val Gln Gly His Pro Gly 2290 2295 2300 Gly Leu Cys Pro Ala Val Ala Thr Tyr Val Ala Ala Cys Gln Ala Ala 2305 2310 2315 2320 Gly Ala Gln Leu Arg Glu Trp Arg Arg Pro Asp Phe Cys Pro Phe Gln 2325 2330 2335 Cys Pro Ala His Ser His Tyr Glu Leu Cys Gly Asp Ser Cys Pro Gly 2340 2345 2350 Ser Cys Pro Ser Leu Ser Ala Pro Glu Gly Cys Glu Ser Ala Cys Arg 2355 2360 2365 Glu Gly Cys Val Cys Asp Ala Gly Phe Val Leu Ser Gly Asp Thr Cys 2370 2375 2380 Val Pro Val Gly Gln Cys Gly Cys Leu His Asp Asp Arg Tyr Tyr Pro 2385 2390 2395 2400 Leu Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys Asp Ser Leu Cys Arg 2405 2410 2415 Cys Arg Glu Gly Gly Glu Val Ser Cys Glu Pro Ser Ser Cys Gly Pro 2420 2425 2430 His Glu Thr Cys Arg Pro Ser Gly Gly Ser Leu Gly Cys Val Ala Val 2435 2440 2445 Gly Ser Thr Thr Cys Gln Ala Ser Gly Asp Pro His Tyr Thr Thr Phe 2450 2455 2460 Asp Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Val Tyr Val Leu Ala 2465 2470 2475 2480 Gln Thr Cys Gly Thr Arg Pro Gly Leu His Arg Phe Ala Val Leu Gln 2485 2490 2495 Glu Asn Val Ala Trp Gly Asn Gly Arg Val Ser Val Thr Arg Val Ile 2500 2505 2510 Thr Val Gln Val Ala Asn Phe Thr Leu Arg Leu Glu Gln Arg Gln Trp 2515 2520 2525 Lys Val Thr Val Asn Gly Val Asp Met Lys Leu Pro Val Val Leu Ala 2530 2535 2540 Asn Gly Gln Ile Arg Ala Ser Gln His Gly Ser Asp Val Val Ile Glu 2545 2550 2555 2560 Thr Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu Val Tyr Tyr Val Arg 2565 2570 2575 Val Thr Val Pro Gly Asn Tyr Tyr Gln Leu Met Cys Gly Leu Cys Gly 2580 2585 2590 Gly Ser 16382 base pairs nucleic acid double linear cDNA not provided CDS 9..16223 8 CTGCAGCC ATG GGT GCC CTA TGG AGC TGG TGG ATA CTC TGG GCT GGA GCA 50 Met Gly Ala Leu Trp Ser Trp Trp Ile Leu Trp Ala Gly Ala 2595 2600 2605 ACC CTC CTG TGG GGA TTG ACC CAG GAG GCT TCA GTG GAC CTC AAG AAC 98 Thr Leu Leu Trp Gly Leu Thr Gln Glu Ala Ser Val Asp Leu Lys Asn 2610 2615 2620 ACT GGC AGA GAG GAA TTC CTC ACA GCC TTC CTG CAG AAC TAT CAG CTG 146 Thr Gly Arg Glu Glu Phe Leu Thr Ala Phe Leu Gln Asn Tyr Gln Leu 2625 2630 2635 2640 GCC TAC AGC AAG GCC TAC CCC CGC CTC CTT ATC TCC AGT CTG TCA GAG 194 Ala Tyr Ser Lys Ala Tyr Pro Arg Leu Leu Ile Ser Ser Leu Ser Glu 2645 2650 2655 AGC CCC GCT TCA GTC TCC ATC CTC AGC CAG GCA GAC AAC ACC TCA AAG 242 Ser Pro Ala Ser Val Ser Ile Leu Ser Gln Ala Asp Asn Thr Ser Lys 2660 2665 2670 AAG GTC ACA GTG AGG CCC GGG GAG TCG GTC ATG GTC AAC ATC AGT GCC 290 Lys Val Thr Val Arg Pro Gly Glu Ser Val Met Val Asn Ile Ser Ala 2675 2680 2685 AAG GCT GAG ATG ATA GGC AGC AAG ATC TTC CAG CAT GCG GTG GTG ATC 338 Lys Ala Glu Met Ile Gly Ser Lys Ile Phe Gln His Ala Val Val Ile 2690 2695 2700 CAT TCT GAC TAT GCC ATC TCT GTG CAG GCA CTA AAT GCC AAG CCT GAC 386 His Ser Asp Tyr Ala Ile Ser Val Gln Ala Leu Asn Ala Lys Pro Asp 2705 2710 2715 2720 ACA GCG GAG CTG ACA CTG CTG CGG CCC ATC CAG GCC CTA GGC ACC GAG 434 Thr Ala Glu Leu Thr Leu Leu Arg Pro Ile Gln Ala Leu Gly Thr Glu 2725 2730 2735 TAT TTT GTG CTC ACA CCC CCC GGC ACC TCA GCC AGG AAT GTC AAG GAG 482 Tyr Phe Val Leu Thr Pro Pro Gly Thr Ser Ala Arg Asn Val Lys Glu 2740 2745 2750 TTT GCC GTG GTG GCC GGT GCC GCA GGT GCC TCG GTC AGT GTC ACG CTG 530 Phe Ala Val Val Ala Gly Ala Ala Gly Ala Ser Val Ser Val Thr Leu 2755 2760 2765 AAG GGG TCA GTG ACA TTC AAT GGC AAG TTC TAT CCA GCA GGC GAT GTC 578 Lys Gly Ser Val Thr Phe Asn Gly Lys Phe Tyr Pro Ala Gly Asp Val 2770 2775 2780 CTA AGA GTG ACT CTA CAG CCC TAC AAT GTG GCC CAG CTA CAG AGC TCA 626 Leu Arg Val Thr Leu Gln Pro Tyr Asn Val Ala Gln Leu Gln Ser Ser 2785 2790 2795 2800 GTG GAT CTC TCG GGG TCA AAG GTC ACA GCT AGT AGC CCC GTG GCT GTC 674 Val Asp Leu Ser Gly Ser Lys Val Thr Ala Ser Ser Pro Val Ala Val 2805 2810 2815 CTC TCT GGC CAC AGC TGT GCG CAG AAA CAT ACG ACC TGC AAC CAT GTG 722 Leu Ser Gly His Ser Cys Ala Gln Lys His Thr Thr Cys Asn His Val 2820 2825 2830 GTT GAG CAG CTG CTA CCC ACG TCT GCC TGG GGC ACC CAC TAT GTA GTA 770 Val Glu Gln Leu Leu Pro Thr Ser Ala Trp Gly Thr His Tyr Val Val 2835 2840 2845 CCC ACG CTG GCC TCC CAA TCT CGC TAT GAT TTG GCC TTC GTT GTG GCC 818 Pro Thr Leu Ala Ser Gln Ser Arg Tyr Asp Leu Ala Phe Val Val Ala 2850 2855 2860 AGC CAG GCC ACA AAG CTG ACC TAC AAC CAT GGG GGT ATC ACT GGC TCC 866 Ser Gln Ala Thr Lys Leu Thr Tyr Asn His Gly Gly Ile Thr Gly Ser 2865 2870 2875 2880 CGT GGG CTC CAG GCA GGT GAT GTG GTA GAG TTT GAG GTC CGG CCA TCC 914 Arg Gly Leu Gln Ala Gly Asp Val Val Glu Phe Glu Val Arg Pro Ser 2885 2890 2895 TGG CCA CTC TAC CTG TCT GCA AAT GTG GGC ATC CAG GTC CTG TTG TTT 962 Trp Pro Leu Tyr Leu Ser Ala Asn Val Gly Ile Gln Val Leu Leu Phe 2900 2905 2910 GGC ACA GGT GCC ATA AGG AAT GAA GTG ACT TAT GAC CCC TAC CTG GTC 1010 Gly Thr Gly Ala Ile Arg Asn Glu Val Thr Tyr Asp Pro Tyr Leu Val 2915 2920 2925 CTG ATC CCA GAT GTG GCG GCC TAC TGC CCA GCC TAT GTG GTC AAG AGT 1058 Leu Ile Pro Asp Val Ala Ala Tyr Cys Pro Ala Tyr Val Val Lys Ser 2930 2935 2940 GTA CCA GGC TGT GAG GGC GTG GCC CTG GTA GTG GCA CAG ACG AAG GCT 1106 Val Pro Gly Cys Glu Gly Val Ala Leu Val Val Ala Gln Thr Lys Ala 2945 2950 2955 2960 ATC AGC GGG CTG ACC ATA GAT GGG CAT GCA GTG GGG GCC AAG CTC ACC 1154 Ile Ser Gly Leu Thr Ile Asp Gly His Ala Val Gly Ala Lys Leu Thr 2965 2970 2975 TGG GAG GCT GTG CCA GGC AGT GAG TTC TCG TAT GCT GAA GTG GAG CTC 1202 Trp Glu Ala Val Pro Gly Ser Glu Phe Ser Tyr Ala Glu Val Glu Leu 2980 2985 2990 GGC ACA GCT GAC ATG ATC CAC ACG GCC GAG GCC ACC ACC AAC TTG GGA 1250 Gly Thr Ala Asp Met Ile His Thr Ala Glu Ala Thr Thr Asn Leu Gly 2995 3000 3005 CTG CTC ACC TTC GGG CTG GCC AAG GCT ATA GGC TAC GCA ACA GCT GCT 1298 Leu Leu Thr Phe Gly Leu Ala Lys Ala Ile Gly Tyr Ala Thr Ala Ala 3010 3015 3020 GAT TGC GGC CGG ACT GTA CTG TCC CCA GTG GAG CCC TCC TGC GAA GGC 1346 Asp Cys Gly Arg Thr Val Leu Ser Pro Val Glu Pro Ser Cys Glu Gly 3025 3030 3035 3040 ATG CAG TGC GCA GCC GGG CAG CGC TGC CAG GTG GTA GGC GGG AAG GCC 1394 Met Gln Cys Ala Ala Gly Gln Arg Cys Gln Val Val Gly Gly Lys Ala 3045 3050 3055 GGG TGT GTG GCG GAG TCC ACC GCT GTC TGC CGC GCC CAG GGC GAC CCC 1442 Gly Cys Val Ala Glu Ser Thr Ala Val Cys Arg Ala Gln Gly Asp Pro 3060 3065 3070 CAT TAC ACC ACC TTC GAC GGC CGT CGC TAC GAC ATG ATG GGC ACC TGT 1490 His Tyr Thr Thr Phe Asp Gly Arg Arg Tyr Asp Met Met Gly Thr Cys 3075 3080 3085 TCG TAC ACG ATG GTG GAG CTG TGC AGC GAG GAC GAC ACC CTG CCC GCC 1538 Ser Tyr Thr Met Val Glu Leu Cys Ser Glu Asp Asp Thr Leu Pro Ala 3090 3095 3100 TTC AGC GTG GAG GCC AAG AAC GAG CAC CGG GGC AGC CGC CGC GTC TCC 1586 Phe Ser Val Glu Ala Lys Asn Glu His Arg Gly Ser Arg Arg Val Ser 3105 3110 3115 3120 TAC GTG GGC CTC GTC ACT GTG CGC GCC TAC AGC CAC TCT GTG TCG CTG 1634 Tyr Val Gly Leu Val Thr Val Arg Ala Tyr Ser His Ser Val Ser Leu 3125 3130 3135 ACC CGC GGT GAA GTT GGC TTC GTC CTG GTT GAC AAC CAG CGC TCG CGC 1682 Thr Arg Gly Glu Val Gly Phe Val Leu Val Asp Asn Gln Arg Ser Arg 3140 3145 3150 CTG CCA GTC TCC CTG AGT GAG GGT CGC CTG CGT GTG TAC CAG AGC GGA 1730 Leu Pro Val Ser Leu Ser Glu Gly Arg Leu Arg Val Tyr Gln Ser Gly 3155 3160 3165 CCA CGG GCC GTG GTG GAG CTG GTC TTT GGG CTG GTG GTC ACT TAT GAC 1778 Pro Arg Ala Val Val Glu Leu Val Phe Gly Leu Val Val Thr Tyr Asp 3170 3175 3180 TGG GAC TGC CAG CTG GCA CTC AGC CTG CCT GCA CGC TTC CAA GAC CAG 1826 Trp Asp Cys Gln Leu Ala Leu Ser Leu Pro Ala Arg Phe Gln Asp Gln 3185 3190 3195 3200 GTG TGC GGG CTG TGT GGC AAC TAT AAT GGT GAC CCA GCA GAC GAC TTC 1874 Val Cys Gly Leu Cys Gly Asn Tyr Asn Gly Asp Pro Ala Asp Asp Phe 3205 3210 3215 CTC ACG CCT GAC GGG GCT CTG GCT CCT GAC GCT GTG GAG TTC GCA AGT 1922 Leu Thr Pro Asp Gly Ala Leu Ala Pro Asp Ala Val Glu Phe Ala Ser 3220 3225 3230 AGC TGG AAG CTG GAT GAT GGG GAC TAC CTG TGT GAG GAT GGC TGC CAG 1970 Ser Trp Lys Leu Asp Asp Gly Asp Tyr Leu Cys Glu Asp Gly Cys Gln 3235 3240 3245 AAC AAC TGT CCC GCC TGC ACC CCA GGC CAG GCC CAA CAC TAT GAG GGC 2018 Asn Asn Cys Pro Ala Cys Thr Pro Gly Gln Ala Gln His Tyr Glu Gly 3250 3255 3260 GAC CGA CTC TGT GGC ATG CTG ACC AAG CTC GAT GGC CCC TTC GCT GTC 2066 Asp Arg Leu Cys Gly Met Leu Thr Lys Leu Asp Gly Pro Phe Ala Val 3265 3270 3275 3280 TGC CAT GAC ACC CTG GAC CCC AGG CCC TTC CTG GAG CAG TGT GTA TAT 2114 Cys His Asp Thr Leu Asp Pro Arg Pro Phe Leu Glu Gln Cys Val Tyr 3285 3290 3295 GAC CTG TGT GTG GTC GGT GGG GAG CGG CTC AGC CTG TGC CGT GGC CTC 2162 Asp Leu Cys Val Val Gly Gly Glu Arg Leu Ser Leu Cys Arg Gly Leu 3300 3305 3310 AGC GCC TAT GCC CAG GCC TGT CTG GAG CTT GGC ATC TCG GTT GGG GAC 2210 Ser Ala Tyr Ala Gln Ala Cys Leu Glu Leu Gly Ile Ser Val Gly Asp 3315 3320 3325 TGG AGA TCA CCA GCC AAC TGC CCC CTG TCC TGC CCT GCC AAC AGC CGC 2258 Trp Arg Ser Pro Ala Asn Cys Pro Leu Ser Cys Pro Ala Asn Ser Arg 3330 3335 3340 TAT GAG CTC TGC GGC CCT GCT TGC CCG ACC TCC TGC AAC GGG GCT GCG 2306 Tyr Glu Leu Cys Gly Pro Ala Cys Pro Thr Ser Cys Asn Gly Ala Ala 3345 3350 3355 3360 GCG CCG TCC AAC TGC TCC GGG CGC CCC TGC GTG GAG GGC TGC GTG TGC 2354 Ala Pro Ser Asn Cys Ser Gly Arg Pro Cys Val Glu Gly Cys Val Cys 3365 3370 3375 CTC CCA GGC TTC GTG GCC AGC GGC GGC GCC TGC GTG CCG GCC TCG TCG 2402 Leu Pro Gly Phe Val Ala Ser Gly Gly Ala Cys Val Pro Ala Ser Ser 3380 3385 3390 TGT GGC TGC ACC TTC CAG GGT CTC CAG CTC GCT CCG GGC CAG GAA GTG 2450 Cys Gly Cys Thr Phe Gln Gly Leu Gln Leu Ala Pro Gly Gln Glu Val 3395 3400 3405 TGG GCG GAC GAG TTG TGC CAA AGG CGC TGC ACC TGC AAC GGC GCC ACC 2498 Trp Ala Asp Glu Leu Cys Gln Arg Arg Cys Thr Cys Asn Gly Ala Thr 3410 3415 3420 CAT CAG GTC ACC TGC CGC GAC AAG CAG AGC TGC CCG GCG GGT GAG CGC 2546 His Gln Val Thr Cys Arg Asp Lys Gln Ser Cys Pro Ala Gly Glu Arg 3425 3430 3435 3440 TGC AGC GTC CAG AAC GGC CTC CTG GGC TGC TAC CCC GAT CGC TTC GGG 2594 Cys Ser Val Gln Asn Gly Leu Leu Gly Cys Tyr Pro Asp Arg Phe Gly 3445 3450 3455 ACC TGC CAG GGG TCC GGG GAC CCA CAC TAT GTG AGC TTC GAC GGC CGG 2642 Thr Cys Gln Gly Ser Gly Asp Pro His Tyr Val Ser Phe Asp Gly Arg 3460 3465 3470 CGC TTC GAC TTC ATG GGC ACC TGC ACG TAC CTG CTG GTC GGC TCA TGC 2690 Arg Phe Asp Phe Met Gly Thr Cys Thr Tyr Leu Leu Val Gly Ser Cys 3475 3480 3485 GGC CAG AAC GCA GCG CTG CCT GCC TTC CGG GTG CTG GTG GAA AAC GAG 2738 Gly Gln Asn Ala Ala Leu Pro Ala Phe Arg Val Leu Val Glu Asn Glu 3490 3495 3500 CAT CGG GGC AGC CAG ACT GTG AGC TAC ACG CGC GCC GTG CGG GTG GAG 2786 His Arg Gly Ser Gln Thr Val Ser Tyr Thr Arg Ala Val Arg Val Glu 3505 3510 3515 3520 GCC CGC GGG GTG AAG GTG GCC GTG CGC CGG GAG TAC CCC GGG CAA GTG 2834 Ala Arg Gly Val Lys Val Ala Val Arg Arg Glu Tyr Pro Gly Gln Val 3525 3530 3535 CTG GTG GAT GAC GTC CTT CAG TAT CTG CCC TTC CAA GCA GCA GAT GGG 2882 Leu Val Asp Asp Val Leu Gln Tyr Leu Pro Phe Gln Ala Ala Asp Gly 3540 3545 3550 CAG GTG CAG GTG TTC CGA CAG GGC AGG GAT GCC GTC GTG CGC ACG GAC 2930 Gln Val Gln Val Phe Arg Gln Gly Arg Asp Ala Val Val Arg Thr Asp 3555 3560 3565 TTT GGC CTG ACT GTC ACT TAT GAC TGG AAT GCA CGA GTG ACT GCC AAG 2978 Phe Gly Leu Thr Val Thr Tyr Asp Trp Asn Ala Arg Val Thr Ala Lys 3570 3575 3580 GTG CCC AGC AGC TAT GCT GAG GCC CTG TGT GGA CTC TGT GGG AAC TTC 3026 Val Pro Ser Ser Tyr Ala Glu Ala Leu Cys Gly Leu Cys Gly Asn Phe 3585 3590 3595 3600 AAC GGG GAC CCA GCT GAT GAC CTG GCT CTG CGG GGT GGG GGT CAA GCT 3074 Asn Gly Asp Pro Ala Asp Asp Leu Ala Leu Arg Gly Gly Gly Gln Ala 3605 3610 3615 GCC AAT GCA CTG GCC TTT GGG AAC AGC TGG CAA GAA GAG ACG AGG CCC 3122 Ala Asn Ala Leu Ala Phe Gly Asn Ser Trp Gln Glu Glu Thr Arg Pro 3620 3625 3630 GGC TGT GGA GCA ACT GAA CCG GGT GAC TGT CCC AAG CTG GAC TCC CTG 3170 Gly Cys Gly Ala Thr Glu Pro Gly Asp Cys Pro Lys Leu Asp Ser Leu 3635 3640 3645 GTG GCC CAG CAG CTG CAG AGC AAG AAT GAG TGT GGA ATC CTT GCC GAC 3218 Val Ala Gln Gln Leu Gln Ser Lys Asn Glu Cys Gly Ile Leu Ala Asp 3650 3655 3660 CCC AAG GGG CCC TTC CGG GAG TGC CAT AGC AAG CTG GAC CCC CAG GGT 3266 Pro Lys Gly Pro Phe Arg Glu Cys His Ser Lys Leu Asp Pro Gln Gly 3665 3670 3675 3680 GCC GTG CGC GAC TGT GTC TAT GAC CGC TGC CTG CTG CCA GGC CAG TCT 3314 Ala Val Arg Asp Cys Val Tyr Asp Arg Cys Leu Leu Pro Gly Gln Ser 3685 3690 3695 GGG CCA CTG TGT GAC GCA CTG GCC ACC TAT GCT GCT GCA TGC CAG GCT 3362 Gly Pro Leu Cys Asp Ala Leu Ala Thr Tyr Ala Ala Ala Cys Gln Ala 3700 3705 3710 GCT GGA GCC ACA GTG CAC CCC TGG AGG AGT GAA GAA CTT TGC CCA CTG 3410 Ala Gly Ala Thr Val His Pro Trp Arg Ser Glu Glu Leu Cys Pro Leu 3715 3720 3725 AGC TGC CCA CCC CAC AGC CAC TAT GAG GCG TGT TCC TAC GGC TGC CCG 3458 Ser Cys Pro Pro His Ser His Tyr Glu Ala Cys Ser Tyr Gly Cys Pro 3730 3735 3740 CTG TCC TGT GGA GAC CTC CCA GTG CCC GGG GGC TGT GGC TCA GAA TGC 3506 Leu Ser Cys Gly Asp Leu Pro Val Pro Gly Gly Cys Gly Ser Glu Cys 3745 3750 3755 3760 CAT GAG GGC TGC GTG TGC GAT GAG GGC TTT GCG CTC AGT GGT GAG TCC 3554 His Glu Gly Cys Val Cys Asp Glu Gly Phe Ala Leu Ser Gly Glu Ser 3765 3770 3775 TGC CTG CCC CTG GCC TCC TGT GGC TGC GTA CAC CAG GGC ACC TAC CAC 3602 Cys Leu Pro Leu Ala Ser Cys Gly Cys Val His Gln Gly Thr Tyr His 3780 3785 3790 CCA CCA GGC CAG ACC TTC TAC CCT GGC CCC GGA TGT GAT TCC CTT TGC 3650 Pro Pro Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys Asp Ser Leu Cys 3795 3800 3805 CAC TGC CAG GAG GGC GGC CTG GTG TCC TGT GAG TCC TCC AGC TGC GGA 3698 His Cys Gln Glu Gly Gly Leu Val Ser Cys Glu Ser Ser Ser Cys Gly 3810 3815 3820 CCG CAC GAG GCC TGC CAG CCA TCC GGT GGC AGC TTG GGC TGT GTG GCC 3746 Pro His Glu Ala Cys Gln Pro Ser Gly Gly Ser Leu Gly Cys Val Ala 3825 3830 3835 3840 GTG GGC TCT AGC ACC TGC CAG GCG TCA GGA GAC CCC CAC TAC ACC ACC 3794 Val Gly Ser Ser Thr Cys Gln Ala Ser Gly Asp Pro His Tyr Thr Thr 3845 3850 3855 TTC GAT GGC CGC CGC TTC GAC TTC ATG GGC ACC TGC GTG TAT GTG CTG 3842 Phe Asp Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Val Tyr Val Leu 3860 3865 3870 GCT CAG ACC TGC GGC ACC CGG CCT GGC CTG CAT CGG TTT GCC GTC CTG 3890 Ala Gln Thr Cys Gly Thr Arg Pro Gly Leu His Arg Phe Ala Val Leu 3875 3880 3885 CAG GAG AAC GTG GCC TGG GGT AAT GGG CGA GTC AGT GTG ACC AGG GTG 3938 Gln Glu Asn Val Ala Trp Gly Asn Gly Arg Val Ser Val Thr Arg Val 3890 3895 3900 ATC ACG GTC CAG GTG GCA AAC TTC ACC CTG CGG CTG GAG CAG AGA CAG 3986 Ile Thr Val Gln Val Ala Asn Phe Thr Leu Arg Leu Glu Gln Arg Gln 3905 3910 3915 3920 TGG AAG GTC ACG GTG AAC GGT GTG GAC ATG AAG CTG CCC GTG GTG CTG 4034 Trp Lys Val Thr Val Asn Gly Val Asp Met Lys Leu Pro Val Val Leu 3925 3930 3935 GCC AAC GGC CAG ATC CGT GCC TCC CAG CAT GGT TCA GAT GTT GTG ATT 4082 Ala Asn Gly Gln Ile Arg Ala Ser Gln His Gly Ser Asp Val Val Ile 3940 3945 3950 GAG ACC GAC TTC GGC CTG CGT GTG GCC TAC GAC CTT GTG TAC TAT GTG 4130 Glu Thr Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu Val Tyr Tyr Val 3955 3960 3965 CGG GTC ACC GTC CCC GGA AAC TAC TAC CAG CAG ATG TGT GGC CTG TGT 4178 Arg Val Thr Val Pro Gly Asn Tyr Tyr Gln Gln Met Cys Gly Leu Cys 3970 3975 3980 GGG AAC TAC AAC GGC GAC CCC AAG GAT GAC TTC CAG AAG CCC AAT GGC 4226 Gly Asn Tyr Asn Gly Asp Pro Lys Asp Asp Phe Gln Lys Pro Asn Gly 3985 3990 3995 4000 TCA CAG GCA GGC AAC GCC AAT GAG TTC GGC AAC TCC TGG GAG GAG GTG 4274 Ser Gln Ala Gly Asn Ala Asn Glu Phe Gly Asn Ser Trp Glu Glu Val 4005 4010 4015 GTG CCC GAC TCT CCC TGC CTG CCG CCC ACC CCT TGC CCG CCG GGG AGC 4322 Val Pro Asp Ser Pro Cys Leu Pro Pro Thr Pro Cys Pro Pro Gly Ser 4020 4025 4030 GAG GAC TGT ATC CCC AGC CAC AAG TGT CCT CCC GAG CTG GAG AAG AAG 4370 Glu Asp Cys Ile Pro Ser His Lys Cys Pro Pro Glu Leu Glu Lys Lys 4035 4040 4045 TAT CAG AAG GAG GAG TTC TGT GGG CTC CTC TCC AGC CCC ACA GGG CCA 4418 Tyr Gln Lys Glu Glu Phe Cys Gly Leu Leu Ser Ser Pro Thr Gly Pro 4050 4055 4060 CTG TCC TCC TGC CAC AAG CTG GTG GAT CCC CAG GGT CCC TTG AAA GAT 4466 Leu Ser Ser Cys His Lys Leu Val Asp Pro Gln Gly Pro Leu Lys Asp 4065 4070 4075 4080 TGC ATC TTT GAT CTC TGC CTG GGT GGT GGG AAC CTG AGC ATT CTC TGC 4514 Cys Ile Phe Asp Leu Cys Leu Gly Gly Gly Asn Leu Ser Ile Leu Cys 4085 4090 4095 AGC AAC ATC CAT GCC TAC GTG AGT GCT TGC CAG GCG GCT GGA GGC CAC 4562 Ser Asn Ile His Ala Tyr Val Ser Ala Cys Gln Ala Ala Gly Gly His 4100 4105 4110 GTG GAG CCC TGG AGG ACT GAA ACT TTC TGT CCC ATG GAG TGC CCT CCG 4610 Val Glu Pro Trp Arg Thr Glu Thr Phe Cys Pro Met Glu Cys Pro Pro 4115 4120 4125 AAC AGT CAC TAC GAG CTC TGT GCG GAC ACC TGC TCC CTG GGC TGC TCA 4658 Asn Ser His Tyr Glu Leu Cys Ala Asp Thr Cys Ser Leu Gly Cys Ser 4130 4135 4140 GCT CTC AGT GCC CCT CCA CAG TGC CAG GAT GGG TGT GCT GAG GGC TGC 4706 Ala Leu Ser Ala Pro Pro Gln Cys Gln Asp Gly Cys Ala Glu Gly Cys 4145 4150 4155 4160 CAG TGT GAC TCC GGC TTC CTC TAC AAT GGC CAA GCC TGC GTG CCC ATC 4754 Gln Cys Asp Ser Gly Phe Leu Tyr Asn Gly Gln Ala Cys Val Pro Ile 4165 4170 4175 CAG CAA TGC GGC TGC TAC CAC AAT GGT GTC TAC TAT GAG CCG GAG CAG 4802 Gln Gln Cys Gly Cys Tyr His Asn Gly Val Tyr Tyr Glu Pro Glu Gln 4180 4185 4190 ACA GTC CTC ATT GAC AAC TGT CGG CAG CAG TGC ACG TGC CAT GCG GGT 4850 Thr Val Leu Ile Asp Asn Cys Arg Gln Gln Cys Thr Cys His Ala Gly 4195 4200 4205 AAA GGC ATG GTG TGC CAG GAA CAC AGC TGC AAG CCG GGG CAG GTG TGC 4898 Lys Gly Met Val Cys Gln Glu His Ser Cys Lys Pro Gly Gln Val Cys 4210 4215 4220 CAG CCC TCC GGA GGC ATC CTG AGC TGC GTC ACC AAA GAC CCG TGC CAC 4946 Gln Pro Ser Gly Gly Ile Leu Ser Cys Val Thr Lys Asp Pro Cys His 4225 4230 4235 4240 GGC GTG ACA TGC CGG CCA CAG GAG ACA TGC AAG GAG CAG GGT GGC CAG 4994 Gly Val Thr Cys Arg Pro Gln Glu Thr Cys Lys Glu Gln Gly Gly Gln 4245 4250 4255 GGC GTG TGC CTG CCC AAC TAT GAG GCC ACG TGC TGG CTG TGG GGC GAC 5042 Gly Val Cys Leu Pro Asn Tyr Glu Ala Thr Cys Trp Leu Trp Gly Asp 4260 4265 4270 CCA CAC TAC CAC TCC TTC GAT GGC CGG AAG TTT GAC TTC CAG GGC ACC 5090 Pro His Tyr His Ser Phe Asp Gly Arg Lys Phe Asp Phe Gln Gly Thr 4275 4280 4285 TGT AAC TAT GTG CTG GCA ACA ACT GGC TGC CCG GGG GTC AGC ACC CAG 5138 Cys Asn Tyr Val Leu Ala Thr Thr Gly Cys Pro Gly Val Ser Thr Gln 4290 4295 4300 GGC CTG ACA CCC TTC ACC GTC ACC ACC AAG AAC CAG AAC CGG GGC AAC 5186 Gly Leu Thr Pro Phe Thr Val Thr Thr Lys Asn Gln Asn Arg Gly Asn 4305 4310 4315 4320 CCT GCT GTG TCC TAC GTG AGA GTC GTC ACC GTG GCT GCC CTC GGC ACC 5234 Pro Ala Val Ser Tyr Val Arg Val Val Thr Val Ala Ala Leu Gly Thr 4325 4330 4335 AAC ATC TCC ATC CAC AAG GAC GAG ATC GGC AAA GTC CGG GTG AAC GGT 5282 Asn Ile Ser Ile His Lys Asp Glu Ile Gly Lys Val Arg Val Asn Gly 4340 4345 4350 GTG CTC ACA GCC TTG CCT GTC TCT GTG GCC GAC GGG CGG ATT TCA GTG 5330 Val Leu Thr Ala Leu Pro Val Ser Val Ala Asp Gly Arg Ile Ser Val 4355 4360 4365 ACC CAG GGT GCA TCG AAG GCA CTG CTG GTG GCT GAC TTT GGA CTG CAA 5378 Thr Gln Gly Ala Ser Lys Ala Leu Leu Val Ala Asp Phe Gly Leu Gln 4370 4375 4380 GTC AGC TAT GAC TGG AAC TGG CGG GTA GAC GTG ACG CTG CCC AGC AGC 5426 Val Ser Tyr Asp Trp Asn Trp Arg Val Asp Val Thr Leu Pro Ser Ser 4385 4390 4395 4400 TAT CAT GGC GCA GTG TGC GGG CTC TGC GGT AAC ATG GAC CGC AAC CCC 5474 Tyr His Gly Ala Val Cys Gly Leu Cys Gly Asn Met Asp Arg Asn Pro 4405 4410 4415 AAC AAT GAC CAG GTC TTC CCT AAT GGC ACA CTG GCT CCC TCC ATA CCC 5522 Asn Asn Asp Gln Val Phe Pro Asn Gly Thr Leu Ala Pro Ser Ile Pro 4420 4425 4430 ATC TGG GGC GGC AGC TGG CGA GCC CCA GGC TGG GAC CCA CTG TGT TGG 5570 Ile Trp Gly Gly Ser Trp Arg Ala Pro Gly Trp Asp Pro Leu Cys Trp 4435 4440 4445 GAC GAA TGT CGG GGG TCC TGC CCA ACG TGC CCT GAG GAC CGG TTG GAG 5618 Asp Glu Cys Arg Gly Ser Cys Pro Thr Cys Pro Glu Asp Arg Leu Glu 4450 4455 4460 CAG TAC GAG GGC CCT GGC TTC TGC GGA CCC CTG GCC CCC GGC ACA GGG 5666 Gln Tyr Glu Gly Pro Gly Phe Cys Gly Pro Leu Ala Pro Gly Thr Gly 4465 4470 4475 4480 GGC CCT TTC ACC ACC TGC CAT GCT CAT GTG CCA CCT GAG AGC TTC TTC 5714 Gly Pro Phe Thr Thr Cys His Ala His Val Pro Pro Glu Ser Phe Phe 4485 4490 4495 AAG GGC TGT GTT CTG GAC GTC TGC ATG GGT GGT GGG GAC CGT GAC ATT 5762 Lys Gly Cys Val Leu Asp Val Cys Met Gly Gly Gly Asp Arg Asp Ile 4500 4505 4510 CTT TGC AAG GCT CTG GCT TCC TAT GTG GCC GCC TGC CAG GCT GCT GGG 5810 Leu Cys Lys Ala Leu Ala Ser Tyr Val Ala Ala Cys Gln Ala Ala Gly 4515 4520 4525 GTT GTC ATC GAA GAC TGG CGG GCA CAG GTT GGC TGT GAG ATC ACC TGC 5858 Val Val Ile Glu Asp Trp Arg Ala Gln Val Gly Cys Glu Ile Thr Cys 4530 4535 4540 CCA GAA AAC AGC CAC TAT GAG GTC TGT GGC CCA CCC TGC CCG GCC AGC 5906 Pro Glu Asn Ser His Tyr Glu Val Cys Gly Pro Pro Cys Pro Ala Ser 4545 4550 4555 4560 TGT CCG TCC CCT GCA CCC CTT ACG ACG CCA GCC GTA TGT GAG GGC CCC 5954 Cys Pro Ser Pro Ala Pro Leu Thr Thr Pro Ala Val Cys Glu Gly Pro 4565 4570 4575 TGT GTG GAG GGC TGC CAG TGC GAC GCG GGT TTC GTG TTA AGT GCT GAC 6002 Cys Val Glu Gly Cys Gln Cys Asp Ala Gly Phe Val Leu Ser Ala Asp 4580 4585 4590 CGC TGT GTT CCC CTC AAC AAC GGC TGC GGC TGC TGG GCC AAT GGC ACC 6050 Arg Cys Val Pro Leu Asn Asn Gly Cys Gly Cys Trp Ala Asn Gly Thr 4595 4600 4605 TAC CAC GAG GCG GGC AGT GAG TTT TGG GCT GAT GGC ACC TGC TCC CAG 6098 Tyr His Glu Ala Gly Ser Glu Phe Trp Ala Asp Gly Thr Cys Ser Gln 4610 4615 4620 TGG TGT CGC TGC GGG CCT GGG GGT GGC TCG CTG GTC TGC ACA CCT GCC 6146 Trp Cys Arg Cys Gly Pro Gly Gly Gly Ser Leu Val Cys Thr Pro Ala 4625 4630 4635 4640 AGC TGT GGG CTG GGT GAA GTG TGT GGC CTC CTG CCA TCC GGC CAG CAC 6194 Ser Cys Gly Leu Gly Glu Val Cys Gly Leu Leu Pro Ser Gly Gln His 4645 4650 4655 GGC TGC CAG CCC GTC AGC ACA GCT GAG TGC CAG GCG TGG GGT GAC CCC 6242 Gly Cys Gln Pro Val Ser Thr Ala Glu Cys Gln Ala Trp Gly Asp Pro 4660 4665 4670 CAT TAC GTC ACT CTG GAT GGG CAC CGA TTC AAT TTC CAA GGC ACC TGC 6290 His Tyr Val Thr Leu Asp Gly His Arg Phe Asn Phe Gln Gly Thr Cys 4675 4680 4685 GAG TAC CTG CTG AGT GCA CCC TGC CAC GGA CCA CCC TTG GGG GCT GAG 6338 Glu Tyr Leu Leu Ser Ala Pro Cys His Gly Pro Pro Leu Gly Ala Glu 4690 4695 4700 AAC TTC ACT GTC ACT GTA GCC AAT GAG CAC CGG GGC AGC CAG GCT GTC 6386 Asn Phe Thr Val Thr Val Ala Asn Glu His Arg Gly Ser Gln Ala Val 4705 4710 4715 4720 AGC TAC ACC CGC AGT GTC ACC CTG CAA ATC TAC AAC CAC AGC CTG ACA 6434 Ser Tyr Thr Arg Ser Val Thr Leu Gln Ile Tyr Asn His Ser Leu Thr 4725 4730 4735 CTG AGT GCC CGC TGG CCC CGG AAG CTA CAG GTG GAC GGC GTG TTC GTC 6482 Leu Ser Ala Arg Trp Pro Arg Lys Leu Gln Val Asp Gly Val Phe Val 4740 4745 4750 ACT CTG CCC TTC CAG CTG GAC TCG CTC CTG CAC GCA CAC CTG AGC GGC 6530 Thr Leu Pro Phe Gln Leu Asp Ser Leu Leu His Ala His Leu Ser Gly 4755 4760 4765 GCC GAC GTG GTG GTG ACC ACA ACC TCA GGG CTC TCG CTG GCT TTC GAC 6578 Ala Asp Val Val Val Thr Thr Thr Ser Gly Leu Ser Leu Ala Phe Asp 4770 4775 4780 GGG GAC AGC TTC GTG CGC CTG CGC GTG CCG GCG GCG TAC GCG GGC TCT 6626 Gly Asp Ser Phe Val Arg Leu Arg Val Pro Ala Ala Tyr Ala Gly Ser 4785 4790 4795 4800 CTC TGT GGC TTA TGC GGG AAC TAC AAC CAG GAC CCC GCA GAC GAC CTG 6674 Leu Cys Gly Leu Cys Gly Asn Tyr Asn Gln Asp Pro Ala Asp Asp Leu 4805 4810 4815 AAG GCG GTG GGC GGG AAG CCC GCC GGA TGG CAG GTG GGC GGC GCC CAG 6722 Lys Ala Val Gly Gly Lys Pro Ala Gly Trp Gln Val Gly Gly Ala Gln 4820 4825 4830 GGC TGC GGG GAA TGT GTG TCC AAG CCA TGC CCG TCG CCG TGC ACC CCA 6770 Gly Cys Gly Glu Cys Val Ser Lys Pro Cys Pro Ser Pro Cys Thr Pro 4835 4840 4845 GAG CAG CAA GAG TCC TTC GGC GGC CCG GAC GCC TGC GGC GTG ATC TCC 6818 Glu Gln Gln Glu Ser Phe Gly Gly Pro Asp Ala Cys Gly Val Ile Ser 4850 4855 4860 GCC ACC GAC GGC CCG CTG GCG CCC TGC CAC GGC CTT GTG CCG CCC GCG 6866 Ala Thr Asp Gly Pro Leu Ala Pro Cys His Gly Leu Val Pro Pro Ala 4865 4870 4875 4880 CAG TAC TTC CAG GGC TGC TTG CTG GAC GCC TGC CAA GTT CAG GGC CAT 6914 Gln Tyr Phe Gln Gly Cys Leu Leu Asp Ala Cys Gln Val Gln Gly His 4885 4890 4895 CCT GGA GGC CTC TGT CCT GCA GTG GCC ACC TAC GTG GCA GCC TGT CAG 6962 Pro Gly Gly Leu Cys Pro Ala Val Ala Thr Tyr Val Ala Ala Cys Gln 4900 4905 4910 GCC GCT GGG GCC CAG CTC CGC GAG TGG AGG CGG CCG GAC TTC TGT CCC 7010 Ala Ala Gly Ala Gln Leu Arg Glu Trp Arg Arg Pro Asp Phe Cys Pro 4915 4920 4925 TTC CAG TGC CCT GCC CAC AGC CAC TAC GAG CTC TGC GGT GAC TCC TGT 7058 Phe Gln Cys Pro Ala His Ser His Tyr Glu Leu Cys Gly Asp Ser Cys 4930 4935 4940 CCT GGG AGC TGC CCG AGC CTG TCG GCA CCC GAG GGC TGT GAG TCG GCC 7106 Pro Gly Ser Cys Pro Ser Leu Ser Ala Pro Glu Gly Cys Glu Ser Ala 4945 4950 4955 4960 TGC CGT GAA GGC TGT GTC TGC GAT GCT GGC TTC GTG CTC AGT GGT GAC 7154 Cys Arg Glu Gly Cys Val Cys Asp Ala Gly Phe Val Leu Ser Gly Asp 4965 4970 4975 ACG TGT GTA CCT GTG GGC CAG TGT GGC TGC CTC CAC GAT GAC CGC TAC 7202 Thr Cys Val Pro Val Gly Gln Cys Gly Cys Leu His Asp Asp Arg Tyr 4980 4985 4990 TAC CCA CTG GGC CAG ACC TTC TAC CCT GGC CCT GGG TGT GAT TCC CTT 7250 Tyr Pro Leu Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys Asp Ser Leu 4995 5000 5005 TGC CGC TGC CGG GAG GGC GGT GAG GTG TCC TGT GAG CCC TCC AGC TGC 7298 Cys Arg Cys Arg Glu Gly Gly Glu Val Ser Cys Glu Pro Ser Ser Cys 5010 5015 5020 GGC CCG CAT GAG ACC TGC CGG CCA TCC GGT GGC AGC TTG GGC TGC GTG 7346 Gly Pro His Glu Thr Cys Arg Pro Ser Gly Gly Ser Leu Gly Cys Val 5025 5030 5035 5040 GCC GTG GGC TCT ACC ACC TGC CAG GCG TCG GGA GAT CCC CAC TAC ACC 7394 Ala Val Gly Ser Thr Thr Cys Gln Ala Ser Gly Asp Pro His Tyr Thr 5045 5050 5055 ACC TTC GAT GGC CGC CGC TTC GAC TTC ATG GGC ACC TGC GTG TAT GTG 7442 Thr Phe Asp Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Val Tyr Val 5060 5065 5070 CTG GCT CAG ACC TGC GGC ACC CGG CCT GGC CTA CAT CGG TTT GCC GTC 7490 Leu Ala Gln Thr Cys Gly Thr Arg Pro Gly Leu His Arg Phe Ala Val 5075 5080 5085 CTG CAG GAG AAC GTG GCC TGG GGT AAT GGG CGA GTC AGT GTG ACC AGG 7538 Leu Gln Glu Asn Val Ala Trp Gly Asn Gly Arg Val Ser Val Thr Arg 5090 5095 5100 GTG ATC ACG GTC CAG GTG GCA AAC TTC ACC CTG CGG CTG GAG CAG AGA 7586 Val Ile Thr Val Gln Val Ala Asn Phe Thr Leu Arg Leu Glu Gln Arg 5105 5110 5115 5120 CAG TGG AAG GTC ACG GTG AAC GGT GTG GAC ATG AAG CTG CCC GTG GTG 7634 Gln Trp Lys Val Thr Val Asn Gly Val Asp Met Lys Leu Pro Val Val 5125 5130 5135 CTG GCC AAC GGC CAG ATC CGT GCC TCC CAG CAT GGT TCA GAT GTT GTG 7682 Leu Ala Asn Gly Gln Ile Arg Ala Ser Gln His Gly Ser Asp Val Val 5140 5145 5150 ATT GAG ACC GAC TTC GGC CTG CGT GTG GCC TAC GAC CTT GTG TAC TAT 7730 Ile Glu Thr Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu Val Tyr Tyr 5155 5160 5165 GTG CGG GTC ACC GTC CCT GGA AAC TAC TAC CAG CTG ATG TGT GGC CTG 7778 Val Arg Val Thr Val Pro Gly Asn Tyr Tyr Gln Leu Met Cys Gly Leu 5170 5175 5180 TGT GGG AAC TAC AAC GGC GAC CCC AAG GAT GAC TTC CAG AAG CCC AAT 7826 Cys Gly Asn Tyr Asn Gly Asp Pro Lys Asp Asp Phe Gln Lys Pro Asn 5185 5190 5195 5200 GGC TCG CAG GCA GGC AAC GCC AAT GAG TTC GGC AAC TCC TGG GAG GAG 7874 Gly Ser Gln Ala Gly Asn Ala Asn Glu Phe Gly Asn Ser Trp Glu Glu 5205 5210 5215 GTG GTG CCC GAC TCT CCC TGC CTG CCG CCG CCC ACC TGC CCG CCG GGG 7922 Val Val Pro Asp Ser Pro Cys Leu Pro Pro Pro Thr Cys Pro Pro Gly 5220 5225 5230 AGC GAG GGC TGT ATC CCC AGC GAG GAG TGT CCT CCC GAG CTG GAG AAG 7970 Ser Glu Gly Cys Ile Pro Ser Glu Glu Cys Pro Pro Glu Leu Glu Lys 5235 5240 5245 AAG TAT CAG AAG GAG GAG TTC TGT GGG CTC CTC TCC AGC CCC ACA GGG 8018 Lys Tyr Gln Lys Glu Glu Phe Cys Gly Leu Leu Ser Ser Pro Thr Gly 5250 5255 5260 CCA CTG TCC TCC TGC CAC AAG CTG GTG GAT CCC CAG GGT CCC TTG AAA 8066 Pro Leu Ser Ser Cys His Lys Leu Val Asp Pro Gln Gly Pro Leu Lys 5265 5270 5275 5280 GAT TGC ATC TTT GAT CTC TGC CTG GGT GGT GGG AAC CTG AGC ATT CTC 8114 Asp Cys Ile Phe Asp Leu Cys Leu Gly Gly Gly Asn Leu Ser Ile Leu 5285 5290 5295 TGC AGC AAC ATC CAT GCC TAC GTG AGT GCT TGC CAG GCG GCT GGA GGC 8162 Cys Ser Asn Ile His Ala Tyr Val Ser Ala Cys Gln Ala Ala Gly Gly 5300 5305 5310 CAC GTG GAG CCC TGG AGG AAT GAA ACT TTC TGT CCC ATG GAA TGC CCT 8210 His Val Glu Pro Trp Arg Asn Glu Thr Phe Cys Pro Met Glu Cys Pro 5315 5320 5325 CAG AAC AGT CAC TAC GAG CTC TGT GCG GAC ACC TGC TCC CTG GGC TGC 8258 Gln Asn Ser His Tyr Glu Leu Cys Ala Asp Thr Cys Ser Leu Gly Cys 5330 5335 5340 TCG GCT CTC AGT GCC CCT CTG CAG TGC CCA GAT GGG TGT GCT GAG GGC 8306 Ser Ala Leu Ser Ala Pro Leu Gln Cys Pro Asp Gly Cys Ala Glu Gly 5345 5350 5355 5360 TGC CAG TGT GAC TCC GGC TTC CTC TAC AAC GGC CAA GCC TGC GTG CCC 8354 Cys Gln Cys Asp Ser Gly Phe Leu Tyr Asn Gly Gln Ala Cys Val Pro 5365 5370 5375 ATC CAG CAA TGT GGC TGC TAC CAC AAT GGT GCC TAC TAT GAG CCG GAG 8402 Ile Gln Gln Cys Gly Cys Tyr His Asn Gly Ala Tyr Tyr Glu Pro Glu 5380 5385 5390 CAG ACA GTC CTC ATT GAC AAC TGT CGG CAG CAG TGC ACG TGC CAT GCG 8450 Gln Thr Val Leu Ile Asp Asn Cys Arg Gln Gln Cys Thr Cys His Ala 5395 5400 5405 GGT AAA GTC GTG GTG TGC CAG GAA CAC AGC TGC AAG CCG GGG CAG GTG 8498 Gly Lys Val Val Val Cys Gln Glu His Ser Cys Lys Pro Gly Gln Val 5410 5415 5420 TGC CAG CCC TCC GGA GGC ATC CTG AGC TGC GTC ACC AAA GAC CCG TGC 8546 Cys Gln Pro Ser Gly Gly Ile Leu Ser Cys Val Thr Lys Asp Pro Cys 5425 5430 5435 5440 CAC GGC GTG ACA TGC CGG CCA CAG GAG ACA TGC AAG GAG CAG GGT GGC 8594 His Gly Val Thr Cys Arg Pro Gln Glu Thr Cys Lys Glu Gln Gly Gly 5445 5450 5455 CAG GGT GTG TGC CTG CCC AAC TAT GAG GCC ACG TGC TGG CTG TGG GGC 8642 Gln Gly Val Cys Leu Pro Asn Tyr Glu Ala Thr Cys Trp Leu Trp Gly 5460 5465 5470 GAC CCA CAC TAC CAC TCC TTC GAT GGC CGG AAG TTT GAC TTC CAG GGC 8690 Asp Pro His Tyr His Ser Phe Asp Gly Arg Lys Phe Asp Phe Gln Gly 5475 5480 5485 ACC TGT AAC TAT GTG CTG GCA ACA ACT GGC TGC CCG GGG GTC AGC ACC 8738 Thr Cys Asn Tyr Val Leu Ala Thr Thr Gly Cys Pro Gly Val Ser Thr 5490 5495 5500 CAG GGC CTG ACA CCC TTC ACC GTC ACC ACC AAG AAC CAG AAC CGG GGC 8786 Gln Gly Leu Thr Pro Phe Thr Val Thr Thr Lys Asn Gln Asn Arg Gly 5505 5510 5515 5520 AAC CCT GCT GTA TCC TAC GTG AGA GTC GTC ACC GTG GCT GCC CTC GGC 8834 Asn Pro Ala Val Ser Tyr Val Arg Val Val Thr Val Ala Ala Leu Gly 5525 5530 5535 ACC AAC ATC TCC ATC CAC AAG GAC GAG ATC GGC AAA GTC CGG GTG AAC 8882 Thr Asn Ile Ser Ile His Lys Asp Glu Ile Gly Lys Val Arg Val Asn 5540 5545 5550 GGT GTG CTC ACA GCC TTG CCT GTC TCC GTG GCC GAC GGG CGG ATT TCA 8930 Gly Val Leu Thr Ala Leu Pro Val Ser Val Ala Asp Gly Arg Ile Ser 5555 5560 5565 GTG GCC CAG GGT GCA TCG AAG GCA CTG CTG GTG GCT GAC TTT GGA CTG 8978 Val Ala Gln Gly Ala Ser Lys Ala Leu Leu Val Ala Asp Phe Gly Leu 5570 5575 5580 CAA GTC AGC TAT GAC TGG AAC TGG CGG GTA GAC GTG ACG CTC CCC AGC 9026 Gln Val Ser Tyr Asp Trp Asn Trp Arg Val Asp Val Thr Leu Pro Ser 5585 5590 5595 5600 AGC TAT CAT GGC GCA GTG TGC GGG CTC TGC GGT AAC ATG GAC CGC AAC 9074 Ser Tyr His Gly Ala Val Cys Gly Leu Cys Gly Asn Met Asp Arg Asn 5605 5610 5615 CCC AAC AAT GAC CAG GTC TTC CCT AAT GGC ACA CTG GCT CCC TCC ATA 9122 Pro Asn Asn Asp Gln Val Phe Pro Asn Gly Thr Leu Ala Pro Ser Ile 5620 5625 5630 CCC ATC TGG GGC GGC AGC TGG CGA GCC CCA GGC TGG GAC CCA CTG TGT 9170 Pro Ile Trp Gly Gly Ser Trp Arg Ala Pro Gly Trp Asp Pro Leu Cys 5635 5640 5645 TGG GAC GAA TGT CGG GGG TCC TGC CCA ACG TGC CCT GAG GAC CGG TTG 9218 Trp Asp Glu Cys Arg Gly Ser Cys Pro Thr Cys Pro Glu Asp Arg Leu 5650 5655 5660 GAG CAG TAC GAG GGC CCT GGC TTC TGC GGA CCC CTG GCC CCC GGC ACA 9266 Glu Gln Tyr Glu Gly Pro Gly Phe Cys Gly Pro Leu Ala Pro Gly Thr 5665 5670 5675 5680 GGG GGC CCT TTC ACC ACC TGC CAT GCT CAT GTG CCA CCT GAG AGC TTC 9314 Gly Gly Pro Phe Thr Thr Cys His Ala His Val Pro Pro Glu Ser Phe 5685 5690 5695 TTC AAG GGC TGT GTT CTG GAC GTC TGC ATG GGT GGT GGG GAC CAT GAC 9362 Phe Lys Gly Cys Val Leu Asp Val Cys Met Gly Gly Gly Asp His Asp 5700 5705 5710 ATT CTT TGC AAG GCT CTG GCT TCC TAC GTG GCC GCC TGC CAG GCC GCT 9410 Ile Leu Cys Lys Ala Leu Ala Ser Tyr Val Ala Ala Cys Gln Ala Ala 5715 5720 5725 GGG GTT GTC ATC GAA GAC TGG CGG GCA CAG GTT GGC TGT GAG ATC ACC 9458 Gly Val Val Ile Glu Asp Trp Arg Ala Gln Val Gly Cys Glu Ile Thr 5730 5735 5740 TGC CCA GAA AAC AGC CAC TAT GAG GTC TGT GGC CCA CCC TGC CCG GCC 9506 Cys Pro Glu Asn Ser His Tyr Glu Val Cys Gly Pro Pro Cys Pro Ala 5745 5750 5755 5760 AGC TGT CCG TCC CCT GCA CCC CTT ACG ACG CCA GCC GTA TGT GAG GGC 9554 Ser Cys Pro Ser Pro Ala Pro Leu Thr Thr Pro Ala Val Cys Glu Gly 5765 5770 5775 CCC TGT GTG GAG GGC TGC CAG TGC GAC GCG GGT TTC GTG TTA AGT GCT 9602 Pro Cys Val Glu Gly Cys Gln Cys Asp Ala Gly Phe Val Leu Ser Ala 5780 5785 5790 GAC CGC TGT GTT CCC CTC AAC AAC GGC TGC GGC TGC TGG GCC AAT GGC 9650 Asp Arg Cys Val Pro Leu Asn Asn Gly Cys Gly Cys Trp Ala Asn Gly 5795 5800 5805 ACC TAC CAC GAG GCG GGC AGT GAG TTT TGG GCT GAT GGC ACC TGC TCC 9698 Thr Tyr His Glu Ala Gly Ser Glu Phe Trp Ala Asp Gly Thr Cys Ser 5810 5815 5820 CAG TGG TGT CGC TGC GGG CCT GGG GGT GGC TCG CTG GTC TGC ACA CCT 9746 Gln Trp Cys Arg Cys Gly Pro Gly Gly Gly Ser Leu Val Cys Thr Pro 5825 5830 5835 5840 GCC AGC TGT GGG CTG GGT GAA GTG TGT GGC CTC CTG CCA TCC GGC CAG 9794 Ala Ser Cys Gly Leu Gly Glu Val Cys Gly Leu Leu Pro Ser Gly Gln 5845 5850 5855 CAC GGC TGC CAG CCC GTC AGC ACA GCT GAG TGC CAG GCG TGG GGT GAC 9842 His Gly Cys Gln Pro Val Ser Thr Ala Glu Cys Gln Ala Trp Gly Asp 5860 5865 5870 CCC CAT TAC GTC ACT CTG GAT GGG CAC CGA TTC GAT TTC CAA GGC ACC 9890 Pro His Tyr Val Thr Leu Asp Gly His Arg Phe Asp Phe Gln Gly Thr 5875 5880 5885 TGC GAG TAC CTG CTG AGT GCA CCC TGC CAC GGA CCA CCC TTG GGG GCT 9938 Cys Glu Tyr Leu Leu Ser Ala Pro Cys His Gly Pro Pro Leu Gly Ala 5890 5895 5900 GAG AAC TTC ACT GTC ACT GTA GCC AAT GAG CAC CGG GGC AGC CAG GCT 9986 Glu Asn Phe Thr Val Thr Val Ala Asn Glu His Arg Gly Ser Gln Ala 5905 5910 5915 5920 GTC AGC TAC ACC CGC AGT GTC ACC CTG CAA ATC TAC AAC CAC AGC CTG 10034 Val Ser Tyr Thr Arg Ser Val Thr Leu Gln Ile Tyr Asn His Ser Leu 5925 5930 5935 ACA CTG AGT GCC CGC TGG CCC CGG AAG CTA CAG GTG GAC GGC GTG TTC 10082 Thr Leu Ser Ala Arg Trp Pro Arg Lys Leu Gln Val Asp Gly Val Phe 5940 5945 5950 GTC ACT CTG CCC TTC CAG CTG GAC TCG CTC CTG CAC GCA CAC CTG AGC 10130 Val Thr Leu Pro Phe Gln Leu Asp Ser Leu Leu His Ala His Leu Ser 5955 5960 5965 GGC GCC GAC GTG GTG GTG ACC ACA ACC TCA GGG CTC TCG CTG GCT TTC 10178 Gly Ala Asp Val Val Val Thr Thr Thr Ser Gly Leu Ser Leu Ala Phe 5970 5975 5980 GAC GGG GAC AGC TTC GTG CGC CTG CGC GTG CCG GCG GCG TAC GCG GGC 10226 Asp Gly Asp Ser Phe Val Arg Leu Arg Val Pro Ala Ala Tyr Ala Gly 5985 5990 5995 6000 TCT CTC TGT GGC TTA TGC GGG AAC TAC AAC CAG GAC CCC GCA GAC GAC 10274 Ser Leu Cys Gly Leu Cys Gly Asn Tyr Asn Gln Asp Pro Ala Asp Asp 6005 6010 6015 CTG AAG GCG GTG GGC GGG AAG CCC GCC GGA TGG CAG GTG GGC GGC GCC 10322 Leu Lys Ala Val Gly Gly Lys Pro Ala Gly Trp Gln Val Gly Gly Ala 6020 6025 6030 CAG GGC TGC GGG GAA TGT GTG TCC AAG CCA TGC CCG TCG CCG TGC ACC 10370 Gln Gly Cys Gly Glu Cys Val Ser Lys Pro Cys Pro Ser Pro Cys Thr 6035 6040 6045 CCA GAG CAG CAA GAG TCC TTC GGC GGC CCG GAC GCC TGC GGC GTG ATC 10418 Pro Glu Gln Gln Glu Ser Phe Gly Gly Pro Asp Ala Cys Gly Val Ile 6050 6055 6060 TCC GCC ACC GAC GGC CCG CTG GCG CCC TGC CAC GGC CTT GTG CCG CCC 10466 Ser Ala Thr Asp Gly Pro Leu Ala Pro Cys His Gly Leu Val Pro Pro 6065 6070 6075 6080 GCG CAG TAC TTC CAG GGC TGC TTG CTG GAC GCC TGC CAA GTT CAG GGC 10514 Ala Gln Tyr Phe Gln Gly Cys Leu Leu Asp Ala Cys Gln Val Gln Gly 6085 6090 6095 CAT CCT GGA GGC CTC TGT CCT GCA GTG GCC ACC TAC GTG GCA GCC TGT 10562 His Pro Gly Gly Leu Cys Pro Ala Val Ala Thr Tyr Val Ala Ala Cys 6100 6105 6110 CAG GCC GCT GGG GCC CAG CTC CGC GAG TGG AGG CGG CCG GAC TTC TGT 10610 Gln Ala Ala Gly Ala Gln Leu Arg Glu Trp Arg Arg Pro Asp Phe Cys 6115 6120 6125 CCC TTC CAG TGC CCT GCC CAC AGC CAC TAC GAG CTC TGC GGT GAC TCC 10658 Pro Phe Gln Cys Pro Ala His Ser His Tyr Glu Leu Cys Gly Asp Ser 6130 6135 6140 TGT CCT GGG AGC TGC CCG AGC CTG TCG GCA CCC GAG GGC TGT GAG TCG 10706 Cys Pro Gly Ser Cys Pro Ser Leu Ser Ala Pro Glu Gly Cys Glu Ser 6145 6150 6155 6160 GCC TGC CGT GAA GGC TGT GTC TGC GAT GCT GGC TTC GTG CTC AGT GGT 10754 Ala Cys Arg Glu Gly Cys Val Cys Asp Ala Gly Phe Val Leu Ser Gly 6165 6170 6175 GAC ACG TGT GTA CCT GTG GGC CAG TGT GGC TGC CTC CAC GAT GAC CGC 10802 Asp Thr Cys Val Pro Val Gly Gln Cys Gly Cys Leu His Asp Asp Arg 6180 6185 6190 TAC TAC CCA CTG GGC CAG ACC TTC TAC CCT GGC CCT GGG TGT GAT TCC 10850 Tyr Tyr Pro Leu Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys Asp Ser 6195 6200 6205 CTT TGC CGC TGC CGG GAG GGC GGT GAG GTG TCC TGT GAG CCC TCC AGC 10898 Leu Cys Arg Cys Arg Glu Gly Gly Glu Val Ser Cys Glu Pro Ser Ser 6210 6215 6220 TGC GGC CCG CAT GAG ACC TGC CGG CCA TCC GGT GGC AGC TTG GGC TGC 10946 Cys Gly Pro His Glu Thr Cys Arg Pro Ser Gly Gly Ser Leu Gly Cys 6225 6230 6235 6240 GTG GCC GTG GGC TCT ACC ACC TGC CAG GCG TCG GGA GAT CCC CAC TAC 10994 Val Ala Val Gly Ser Thr Thr Cys Gln Ala Ser Gly Asp Pro His Tyr 6245 6250 6255 ACC ACC TTC GAT GGC CGC CGC TTC GAC TTC ATG GGC ACC TGC GTG TAT 11042 Thr Thr Phe Asp Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Val Tyr 6260 6265 6270 GTG CTG GCT CAG ACC TGC GGC ACC CGG CCT GGC CTA CAT CGG TTT GCC 11090 Val Leu Ala Gln Thr Cys Gly Thr Arg Pro Gly Leu His Arg Phe Ala 6275 6280 6285 GTC CTG CAG GAG AAC GTG GCC TGG GGT AAT GGG CGA GTC AGT GTG ACC 11138 Val Leu Gln Glu Asn Val Ala Trp Gly Asn Gly Arg Val Ser Val Thr 6290 6295 6300 AGG GTG ATC ACG GTC CAG GTG GCA AAC TTC ACC CTG CGG CTG GAG CAG 11186 Arg Val Ile Thr Val Gln Val Ala Asn Phe Thr Leu Arg Leu Glu Gln 6305 6310 6315 6320 AGA CAG TGG AAG GTC ACG GTG AAC GGT GTG GAC ATG AAG CTG CCC GTG 11234 Arg Gln Trp Lys Val Thr Val Asn Gly Val Asp Met Lys Leu Pro Val 6325 6330 6335 GTG CTG GCC AAC GGC CAG ATC CGT GCC TCC CAG CAT GGT TCA GAT GTT 11282 Val Leu Ala Asn Gly Gln Ile Arg Ala Ser Gln His Gly Ser Asp Val 6340 6345 6350 GTG ATT GAG ACC GAC TTC GGC CTG CGT GTG GCC TAC GAC CTT GTG TAC 11330 Val Ile Glu Thr Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu Val Tyr 6355 6360 6365 TAT GTG CGG GTC ACC GTC CCT GGA AAC TAC TAC CAG CTG ATG TGT GGC 11378 Tyr Val Arg Val Thr Val Pro Gly Asn Tyr Tyr Gln Leu Met Cys Gly 6370 6375 6380 CTG TGT GGG AAC TAC AAC GGC GAC CCC AAG GAT GAC TTC CAG AAG CCC 11426 Leu Cys Gly Asn Tyr Asn Gly Asp Pro Lys Asp Asp Phe Gln Lys Pro 6385 6390 6395 6400 AAT GGC TCG CAG GCA GGC AAC GCC AAT GAG TTC GGC AAC TCC TGG GAG 11474 Asn Gly Ser Gln Ala Gly Asn Ala Asn Glu Phe Gly Asn Ser Trp Glu 6405 6410 6415 GAG GTG GTG CCC GAC TCT CCC TGC CTG CCG CCG CCC ACC TGC CCG CCG 11522 Glu Val Val Pro Asp Ser Pro Cys Leu Pro Pro Pro Thr Cys Pro Pro 6420 6425 6430 GGG AGC GAG GGC TGT ATC CCC AGC GAG GAG TGT CCT CCC GAG CTG GAG 11570 Gly Ser Glu Gly Cys Ile Pro Ser Glu Glu Cys Pro Pro Glu Leu Glu 6435 6440 6445 AAG AAG TAT CAG AAG GAG GAG TTC TGT GGG CTC CTC TCC AGC CCC ACA 11618 Lys Lys Tyr Gln Lys Glu Glu Phe Cys Gly Leu Leu Ser Ser Pro Thr 6450 6455 6460 GGG CCA CTG TCC TCC TGC CAC AAG CTG GTG GAT CCC CAG GGT CCC TTG 11666 Gly Pro Leu Ser Ser Cys His Lys Leu Val Asp Pro Gln Gly Pro Leu 6465 6470 6475 6480 AAA GAT TGC ATC TTT GAT CTC TGC CTG GGT GGT GGG AAC CTG AGC ATT 11714 Lys Asp Cys Ile Phe Asp Leu Cys Leu Gly Gly Gly Asn Leu Ser Ile 6485 6490 6495 CTC TGC AGC AAC ATC CAT GCC TAC GTG AGT GCT TGC CAG GCG GCT GGA 11762 Leu Cys Ser Asn Ile His Ala Tyr Val Ser Ala Cys Gln Ala Ala Gly 6500 6505 6510 GGC CAC GTG GAG CCC TGG AGG AAT GAA ACT TTC TGT CCC ATG GAA TGC 11810 Gly His Val Glu Pro Trp Arg Asn Glu Thr Phe Cys Pro Met Glu Cys 6515 6520 6525 CCT CAG AAC AGT CAC TAC GAG CTC TGT GCG GAC ACC TGC TCC CTG GGC 11858 Pro Gln Asn Ser His Tyr Glu Leu Cys Ala Asp Thr Cys Ser Leu Gly 6530 6535 6540 TGC TCG GCT CTC AGT GCC CCT CTG CAG TGC CCA GAT GGG TGT GCT GAG 11906 Cys Ser Ala Leu Ser Ala Pro Leu Gln Cys Pro Asp Gly Cys Ala Glu 6545 6550 6555 6560 GGC TGC CAG TGT GAC TCC GGC TTC CTC TAC AAC GGC CAA GCC TGC GTG 11954 Gly Cys Gln Cys Asp Ser Gly Phe Leu Tyr Asn Gly Gln Ala Cys Val 6565 6570 6575 CCC ATC CAG CAA TGT GGC TGC TAC CAC AAT GGT GTC TAC TAT GAG CCG 12002 Pro Ile Gln Gln Cys Gly Cys Tyr His Asn Gly Val Tyr Tyr Glu Pro 6580 6585 6590 GAG CAG ACA GTC CTC ATT GAC AAC TGT CGG CAG CAG TGC ACG TGC CAT 12050 Glu Gln Thr Val Leu Ile Asp Asn Cys Arg Gln Gln Cys Thr Cys His 6595 6600 6605 GTG GGT AAA GTC GTG GTG TGC CAG GAA CAC AGC TGC AAG CCG GGG CAG 12098 Val Gly Lys Val Val Val Cys Gln Glu His Ser Cys Lys Pro Gly Gln 6610 6615 6620 GTG TGC CAG CCC TCC GGA GGC ATC CTG AGC TGC GTC AAC AAA GAC CCG 12146 Val Cys Gln Pro Ser Gly Gly Ile Leu Ser Cys Val Asn Lys Asp Pro 6625 6630 6635 6640 TGC CAC GGC GTG ACA TGC CGG CCA CAG GAG ACA TGC AAG GAG CAG GGT 12194 Cys His Gly Val Thr Cys Arg Pro Gln Glu Thr Cys Lys Glu Gln Gly 6645 6650 6655 GGC CAG GGT GTG TGC CTG CCC AAC TAT GAG GCC ACG TGC TGG CTG TGG 12242 Gly Gln Gly Val Cys Leu Pro Asn Tyr Glu Ala Thr Cys Trp Leu Trp 6660 6665 6670 GGC GAC CCA CAC TAC CAC TCC TTC GAT GGC CGG AAG TTT GAC TTC CAG 12290 Gly Asp Pro His Tyr His Ser Phe Asp Gly Arg Lys Phe Asp Phe Gln 6675 6680 6685 GGC ACC TGT AAC TAT GTG CTG GCA ACA ACT GGC TGC CCG GGG GTC AGC 12338 Gly Thr Cys Asn Tyr Val Leu Ala Thr Thr Gly Cys Pro Gly Val Ser 6690 6695 6700 ACC CAG GGC CTG ACA CCC TTC ACC GTC ACC ACC AAG AAC CAG AAC CGG 12386 Thr Gln Gly Leu Thr Pro Phe Thr Val Thr Thr Lys Asn Gln Asn Arg 6705 6710 6715 6720 GGC AAC CCT GCT GTA TCC TAC GTG AGA GTC GTC ACC GTG GCT GCC CTC 12434 Gly Asn Pro Ala Val Ser Tyr Val Arg Val Val Thr Val Ala Ala Leu 6725 6730 6735 GGC ACC AAC ATC TCC ATC CAC AAG GAC GAG ATC GGC AAA GTC CGG GTG 12482 Gly Thr Asn Ile Ser Ile His Lys Asp Glu Ile Gly Lys Val Arg Val 6740 6745 6750 AAC GGT GTG CTC ACA GCC TTG CCT GTC TCC GTG GCC GAC GGG CGG ATT 12530 Asn Gly Val Leu Thr Ala Leu Pro Val Ser Val Ala Asp Gly Arg Ile 6755 6760 6765 TCA GTG GCC CAG GGT GCA TCG AAG GCA CTG CTG GTG GCT GAC TTT GGA 12578 Ser Val Ala Gln Gly Ala Ser Lys Ala Leu Leu Val Ala Asp Phe Gly 6770 6775 6780 CTG CAA GTC AGC TAT GAC TGG AAC TGG CGG GTA GAC GTG ACG CTC CCC 12626 Leu Gln Val Ser Tyr Asp Trp Asn Trp Arg Val Asp Val Thr Leu Pro 6785 6790 6795 6800 AGC AGC TAT CAT GGC GCA GTG TGC GGG CTC TGC GGT AAC ATG GAC CGC 12674 Ser Ser Tyr His Gly Ala Val Cys Gly Leu Cys Gly Asn Met Asp Arg 6805 6810 6815 AAC CCC AAC AAT GAC CAG GTC TTC CCT AAT GGC ACA CTG GCT CCC TCC 12722 Asn Pro Asn Asn Asp Gln Val Phe Pro Asn Gly Thr Leu Ala Pro Ser 6820 6825 6830 ATA CCC ATC TGG GGC GGC AGC TGG CGA GCC CCA GGC TGG GAC CCA CTG 12770 Ile Pro Ile Trp Gly Gly Ser Trp Arg Ala Pro Gly Trp Asp Pro Leu 6835 6840 6845 TGT TGG GAC GAA TGT CGG GGG TCC TGC CCA ACG TGC CCT GAG GAC CGG 12818 Cys Trp Asp Glu Cys Arg Gly Ser Cys Pro Thr Cys Pro Glu Asp Arg 6850 6855 6860 TTG GAG CAG TAC GAG GGG CCT GGC TTC TGC GGA CCC CTG GCA TCT GGC 12866 Leu Glu Gln Tyr Glu Gly Pro Gly Phe Cys Gly Pro Leu Ala Ser Gly 6865 6870 6875 6880 ACA GGG GGC CCC TTC ACC ACC TGC CAT GCT CAT GTG CCA CCT GAG AGC 12914 Thr Gly Gly Pro Phe Thr Thr Cys His Ala His Val Pro Pro Glu Ser 6885 6890 6895 TTC TTC AAG GGC TGT GTT CTG GAC GTC TGC ATG GGT GGT GGG GAC CAT 12962 Phe Phe Lys Gly Cys Val Leu Asp Val Cys Met Gly Gly Gly Asp His 6900 6905 6910 GAC ATT CTT TGC AAG GCT CTG GCT TCC TAC GTG GCC GCC TGC CAG GCC 13010 Asp Ile Leu Cys Lys Ala Leu Ala Ser Tyr Val Ala Ala Cys Gln Ala 6915 6920 6925 GCT GGG GTT GTC ATC GAA GAC TGG CGG GCA CAG GTT GGC TGT GAG ATC 13058 Ala Gly Val Val Ile Glu Asp Trp Arg Ala Gln Val Gly Cys Glu Ile 6930 6935 6940 ACC TGC CCA GAA AAC AGC CAC TAT GAG GTC TGT GGC CCA CCC TGC CCG 13106 Thr Cys Pro Glu Asn Ser His Tyr Glu Val Cys Gly Pro Pro Cys Pro 6945 6950 6955 6960 GCC AGC TGT CCG TCC CCT GCA CCC CTT ACG ACG CCA GCC GTA TGT GAG 13154 Ala Ser Cys Pro Ser Pro Ala Pro Leu Thr Thr Pro Ala Val Cys Glu 6965 6970 6975 GGC CCC TGT GTG GAG GGC TGC CAG TGC GAC GCG GGT TTC GTG TTA AGT 13202 Gly Pro Cys Val Glu Gly Cys Gln Cys Asp Ala Gly Phe Val Leu Ser 6980 6985 6990 GCT GAC CGC TGT GTT CCC CTC AAC AAC GGC TGC GGC TGC TGG GCC AAT 13250 Ala Asp Arg Cys Val Pro Leu Asn Asn Gly Cys Gly Cys Trp Ala Asn 6995 7000 7005 GGC ACC TAC CAC GAG GCG GGC AGT GAG TTT TGG GCT GAT GGC ACC TGC 13298 Gly Thr Tyr His Glu Ala Gly Ser Glu Phe Trp Ala Asp Gly Thr Cys 7010 7015 7020 TCC CAG TGG TGT CGC TGC GGG CCT GGG GGT GGC TCG CTG GTC TGC ACA 13346 Ser Gln Trp Cys Arg Cys Gly Pro Gly Gly Gly Ser Leu Val Cys Thr 7025 7030 7035 7040 CCT GCC AGC TGT GGG CTG GGT GAA GTG TGT GGC CTC CTG CCA TCC GGC 13394 Pro Ala Ser Cys Gly Leu Gly Glu Val Cys Gly Leu Leu Pro Ser Gly 7045 7050 7055 CAG CAC AGC TGC CAG CCC GTC AGC ACA GCT GAG TGC CAG GCG TGG GGT 13442 Gln His Ser Cys Gln Pro Val Ser Thr Ala Glu Cys Gln Ala Trp Gly 7060 7065 7070 GAC CCC CAT TAC GTC ACT CTG GAT GGG CAC CGA TTC GAT TTC CAA GGC 13490 Asp Pro His Tyr Val Thr Leu Asp Gly His Arg Phe Asp Phe Gln Gly 7075 7080 7085 ACC TGC GAG TAC CTG CTG AGT GCA CCC TGC CAC GGA CCA CCC TTG GGG 13538 Thr Cys Glu Tyr Leu Leu Ser Ala Pro Cys His Gly Pro Pro Leu Gly 7090 7095 7100 GCT GAG AAC TTC ACT GTC ACT GTA GCC AAT GAG CAC CGG GGC AGC CAG 13586 Ala Glu Asn Phe Thr Val Thr Val Ala Asn Glu His Arg Gly Ser Gln 7105 7110 7115 7120 GCT GTC AGC TAC ACC CGC AGT GTC ACC CTG CAA ATC TAC AAC CAC AGC 13634 Ala Val Ser Tyr Thr Arg Ser Val Thr Leu Gln Ile Tyr Asn His Ser 7125 7130 7135 CTG ACA CTG AGT GCC CGC TGG CCC CGG AAG CTA CAG GTC GAC GGC GTG 13682 Leu Thr Leu Ser Ala Arg Trp Pro Arg Lys Leu Gln Val Asp Gly Val 7140 7145 7150 TTC GTG GCT CTG CCT TTC CAG CTG GAC TCG CTC CTG CAC GCA CAC CTG 13730 Phe Val Ala Leu Pro Phe Gln Leu Asp Ser Leu Leu His Ala His Leu 7155 7160 7165 AGC GGC GCC GAC GTG GTG GTG ACC ACA ACC TCA GGG CTC TCG CTG GCT 13778 Ser Gly Ala Asp Val Val Val Thr Thr Thr Ser Gly Leu Ser Leu Ala 7170 7175 7180 TTC GAT GGG GAC AGC TTC GTG CGC CTG CGC GTG CCG GCG GCG TAC GCG 13826 Phe Asp Gly Asp Ser Phe Val Arg Leu Arg Val Pro Ala Ala Tyr Ala 7185 7190 7195 7200 GCC TCT CTC TGT GGC TTA TGC GGG AAC TAC AAC CAG GAC CCC GCA GAC 13874 Ala Ser Leu Cys Gly Leu Cys Gly Asn Tyr Asn Gln Asp Pro Ala Asp 7205 7210 7215 GAC CTG AAG GCT GTG GGC GGG AAG CCC GCT GGA TGG CAG GTG GGC GGG 13922 Asp Leu Lys Ala Val Gly Gly Lys Pro Ala Gly Trp Gln Val Gly Gly 7220 7225 7230 GCC CAG GGC TGC GGG GAA TGT GTG TCC AAG CCA TGC CCG TCG CCG TGC 13970 Ala Gln Gly Cys Gly Glu Cys Val Ser Lys Pro Cys Pro Ser Pro Cys 7235 7240 7245 ACC CCA GAG CAG CAG GAG TCC TTC GGC GGC CCG GAC GCC TGC GGC GTG 14018 Thr Pro Glu Gln Gln Glu Ser Phe Gly Gly Pro Asp Ala Cys Gly Val 7250 7255 7260 ATC TCC GCC ACC GAC GGC CCG CTG GCA CCC TGC CAC GGC CTT GTG CCG 14066 Ile Ser Ala Thr Asp Gly Pro Leu Ala Pro Cys His Gly Leu Val Pro 7265 7270 7275 7280 CCC GCG CAG TAC TTC CAG GGC TGC TTG CTG GAC GCC TGC CAA GTT CAG 14114 Pro Ala Gln Tyr Phe Gln Gly Cys Leu Leu Asp Ala Cys Gln Val Gln 7285 7290 7295 GGC CAT CCT GGA GGC CTC TGT CCT GCA GTG GCT ACC TAC GTG GCA GCC 14162 Gly His Pro Gly Gly Leu Cys Pro Ala Val Ala Thr Tyr Val Ala Ala 7300 7305 7310 TGT CAG GCC GCT GGG GCC CAG CTC GGC GAG TGG AGG CGG CCG GAC TTC 14210 Cys Gln Ala Ala Gly Ala Gln Leu Gly Glu Trp Arg Arg Pro Asp Phe 7315 7320 7325 TGT CCC TTG CAG TGC CCT GCC CAC AGC CAC TAT GAG CTC TGC GGT GAC 14258 Cys Pro Leu Gln Cys Pro Ala His Ser His Tyr Glu Leu Cys Gly Asp 7330 7335 7340 TCC TGC CCT GTG AGC TGC CCG AGC CTC TCA GCA CCC GAG GGC TGT GAG 14306 Ser Cys Pro Val Ser Cys Pro Ser Leu Ser Ala Pro Glu Gly Cys Glu 7345 7350 7355 7360 TCG GCC TGC CGT GAA GGC TGT GTC TGC GAT GCT GGC TTC GTA CTC AGT 14354 Ser Ala Cys Arg Glu Gly Cys Val Cys Asp Ala Gly Phe Val Leu Ser 7365 7370 7375 GGT GAC ACC TGC GTA CCC GTG GGC CAG TGT GGC TGC CTC CAT GAT GGC 14402 Gly Asp Thr Cys Val Pro Val Gly Gln Cys Gly Cys Leu His Asp Gly 7380 7385 7390 CGC TAC TAC CCA CTG GGC GAG GTC TTC TAC CCG GGC CCT GAG TGT GAG 14450 Arg Tyr Tyr Pro Leu Gly Glu Val Phe Tyr Pro Gly Pro Glu Cys Glu 7395 7400 7405 CGA CGC TGT GAG TGT GGG CCA GGT GGC CAT GTC ACC TGC CAG GAG GGC 14498 Arg Arg Cys Glu Cys Gly Pro Gly Gly His Val Thr Cys Gln Glu Gly 7410 7415 7420 GCA GCC TGT GGG CCC CAT GAG GAG TGC CGG TTA GAG GAT GGT GTC CAG 14546 Ala Ala Cys Gly Pro His Glu Glu Cys Arg Leu Glu Asp Gly Val Gln 7425 7430 7435 7440 GCC TGT CAT GCC ACA GGC TGT GGC CGC TGC CTG GCC AAC GGG GGC ATC 14594 Ala Cys His Ala Thr Gly Cys Gly Arg Cys Leu Ala Asn Gly Gly Ile 7445 7450 7455 CAC TAC ATC ACC CTT GAT GGC CGT GTC TAC GAC CTG CAT GGC TCC TGC 14642 His Tyr Ile Thr Leu Asp Gly Arg Val Tyr Asp Leu His Gly Ser Cys 7460 7465 7470 TCC TAT GTC TTG GCC CAA GTC TGC CAC CCA AAG CCT GGG GAC GAG GAC 14690 Ser Tyr Val Leu Ala Gln Val Cys His Pro Lys Pro Gly Asp Glu Asp 7475 7480 7485 TTT TCC ATC GTG CTT GAG AAG AAT GCA GCT GGA CAT CTC CAA CGC CTC 14738 Phe Ser Ile Val Leu Glu Lys Asn Ala Ala Gly His Leu Gln Arg Leu 7490 7495 7500 CTG GTT ACT GTG GCT GGC CAG GTT GTG AGC CTA GCT CAG GGG CAG CAG 14786 Leu Val Thr Val Ala Gly Gln Val Val Ser Leu Ala Gln Gly Gln Gln 7505 7510 7515 7520 GTC ACC GTG GAC GGC GAG GCT GTG GCC CTG CCT GTG GCT GTG GGC CGC 14834 Val Thr Val Asp Gly Glu Ala Val Ala Leu Pro Val Ala Val Gly Arg 7525 7530 7535 GTG CGG GTG ACC GCC GAG GGC CGA AAC ATG GTT CTG CAG ACG ACC AAG 14882 Val Arg Val Thr Ala Glu Gly Arg Asn Met Val Leu Gln Thr Thr Lys 7540 7545 7550 GGG CTG CGG CTT CTC TTT GAT GGC GAT GCC CAC CTC CTC ATG TCC ATC 14930 Gly Leu Arg Leu Leu Phe Asp Gly Asp Ala His Leu Leu Met Ser Ile 7555 7560 7565 CCC AGC CCC TTC CGT GGA CGG CTC TGT GGC CTC TGT GGG AAC TTC AAT 14978 Pro Ser Pro Phe Arg Gly Arg Leu Cys Gly Leu Cys Gly Asn Phe Asn 7570 7575 7580 GGC AAC TGG AGT GAC GAC TTT GTC CTG CCC AAT GGC TCA GCA GCG TCC 15026 Gly Asn Trp Ser Asp Asp Phe Val Leu Pro Asn Gly Ser Ala Ala Ser 7585 7590 7595 7600 AGT GTG GAG ACC TTC GGG GCT GCA TGG CGG GTG CCC GGC TCC TCC AAG 15074 Ser Val Glu Thr Phe Gly Ala Ala Trp Arg Val Pro Gly Ser Ser Lys 7605 7610 7615 GGC TGT GGC GAG GGC TGC GGG CCC CAA GGC TGC CCA GTG TGC TTG GCA 15122 Gly Cys Gly Glu Gly Cys Gly Pro Gln Gly Cys Pro Val Cys Leu Ala 7620 7625 7630 GAG GAG ACT GCA CCC TAT GAG AGC AAC GAG GCC TGC GGG CAG CTC CGG 15170 Glu Glu Thr Ala Pro Tyr Glu Ser Asn Glu Ala Cys Gly Gln Leu Arg 7635 7640 7645 AAC CCC CAG GGC CCC TTC GCG ACC TGC CAG GCG GTG CTG AGT CCC TCT 15218 Asn Pro Gln Gly Pro Phe Ala Thr Cys Gln Ala Val Leu Ser Pro Ser 7650 7655 7660 GAG TAC TTC CGC CAA TGC GTA TAC GAC CTG TGC GCG CAA AAG GGT GAC 15266 Glu Tyr Phe Arg Gln Cys Val Tyr Asp Leu Cys Ala Gln Lys Gly Asp 7665 7670 7675 7680 AAA GCC TTC CTG TGC CGC AGC CTG GCA GCC TAC ACG GCG GCC TGT CAG 15314 Lys Ala Phe Leu Cys Arg Ser Leu Ala Ala Tyr Thr Ala Ala Cys Gln 7685 7690 7695 GCA GCT GGC GTG GCC GTG AAG CCC TGG AGG ACA GAC AGC TTC TGC CCG 15362 Ala Ala Gly Val Ala Val Lys Pro Trp Arg Thr Asp Ser Phe Cys Pro 7700 7705 7710 CTC CAT TGC CCC GCC CAC AGC CAC TAC TCC ATC TGC ACT CGC ACC TGC 15410 Leu His Cys Pro Ala His Ser His Tyr Ser Ile Cys Thr Arg Thr Cys 7715 7720 7725 CAG GGA TCC TGT GCG GCT CTC TCC GGC CTC ACG GGC TGC ACC ACC CGC 15458 Gln Gly Ser Cys Ala Ala Leu Ser Gly Leu Thr Gly Cys Thr Thr Arg 7730 7735 7740 TGT TTT GAG GGC TGT GAG TGC GAC GAC CGC TTC CTG CTT TCC CAG GGT 15506 Cys Phe Glu Gly Cys Glu Cys Asp Asp Arg Phe Leu Leu Ser Gln Gly 7745 7750 7755 7760 GTC TGC ATC CCT GTC CAA GAT TGT GGC TGC ACC CAT AAT GGC CGA TAC 15554 Val Cys Ile Pro Val Gln Asp Cys Gly Cys Thr His Asn Gly Arg Tyr 7765 7770 7775 TTG CCG GTA AAC TCC TCC CTG CTG ACC TCA GAC TGC AGC GAG CGC TGT 15602 Leu Pro Val Asn Ser Ser Leu Leu Thr Ser Asp Cys Ser Glu Arg Cys 7780 7785 7790 TCC TGT TCC TCA AGC TCT GGC CTG ACA TGC CAG GCC GCT GGC TGC CCA 15650 Ser Cys Ser Ser Ser Ser Gly Leu Thr Cys Gln Ala Ala Gly Cys Pro 7795 7800 7805 CCA GGC CGT GTA TGT GAG GTC AAG GCT GAA GCC CGG AAC TGC TGG GCC 15698 Pro Gly Arg Val Cys Glu Val Lys Ala Glu Ala Arg Asn Cys Trp Ala 7810 7815 7820 ACC CGT GGT CTC TGT GTC CTG TCT GTG GGT GCC AAC CTC ACC ACC TTT 15746 Thr Arg Gly Leu Cys Val Leu Ser Val Gly Ala Asn Leu Thr Thr Phe 7825 7830 7835 7840 GAT GGG GCC CGT GGT GCC ACC ACC TCT CCT GGT GTC TAT GAG CTC TCT 15794 Asp Gly Ala Arg Gly Ala Thr Thr Ser Pro Gly Val Tyr Glu Leu Ser 7845 7850 7855 TCC CGC TGC CCA GGA CTA CAG AAT ACC ATC CCC TGG TAC CGT GTA GTT 15842 Ser Arg Cys Pro Gly Leu Gln Asn Thr Ile Pro Trp Tyr Arg Val Val 7860 7865 7870 GCC GAA GTC CAG ATC TGC CAT GGC AAA ACG GAG GCT GTG GGC CAG GTC 15890 Ala Glu Val Gln Ile Cys His Gly Lys Thr Glu Ala Val Gly Gln Val 7875 7880 7885 CAC ATC TTC TTC CAG GAT GGG ATG GTG ACG TTG ACT CCA AAC AAG GGT 15938 His Ile Phe Phe Gln Asp Gly Met Val Thr Leu Thr Pro Asn Lys Gly 7890 7895 7900 GTG TGG GTG AAT GGT CTC CGA GTG GAT CTC CCA GCT GAG AAG TTA GCA 15986 Val Trp Val Asn Gly Leu Arg Val Asp Leu Pro Ala Glu Lys Leu Ala 7905 7910 7915 7920 TCT GTG TCC GTG AGT CGT ACA CCT GAT GGC TCC CTG CTA GTC CGC CAG 16034 Ser Val Ser Val Ser Arg Thr Pro Asp Gly Ser Leu Leu Val Arg Gln 7925 7930 7935 AAG GCA GGG GTC CAG GTG TGG CTT GGA GCC AAT GGG AAG GTG GCT GTG 16082 Lys Ala Gly Val Gln Val Trp Leu Gly Ala Asn Gly Lys Val Ala Val 7940 7945 7950 ATT GTC AGC AAT GAC CAT GCT GGG AAA CTG TGT GGG GCC TGT GGA AAC 16130 Ile Val Ser Asn Asp His Ala Gly Lys Leu Cys Gly Ala Cys Gly Asn 7955 7960 7965 TTT GAC GGG GAC CAG ACC AAT GAT TGG CAT GAC TCC CAG GAG AAG CCA 16178 Phe Asp Gly Asp Gln Thr Asn Asp Trp His Asp Ser Gln Glu Lys Pro 7970 7975 7980 GCG ATG GAG AAA TGG AGA GCG CAG GAC TTC TCC CCA TGT TAT GGC 16223 Ala Met Glu Lys Trp Arg Ala Gln Asp Phe Ser Pro Cys Tyr Gly 7985 7990 7995 TGATCAGTCA TCCACCAGGA ACGAAGATTT CCTGAAGAAG ACCTGGTCCC TCTGGAGGTT 16283 GCGGTGGCTG AAGGATGCAT CATGTGCTCC TACCCTGCTC TACCGCTTTT CTGGGTCACA 16343 GAGGCCAAAT GTGAGAGCAT TGAATAAATA TCTTAAGCT 16382 5405 amino acids amino acid linear protein not provided 9 Met Gly Ala Leu Trp Ser Trp Trp Ile Leu Trp Ala Gly Ala Thr Leu 1 5 10 15 Leu Trp Gly Leu Thr Gln Glu Ala Ser Val Asp Leu Lys Asn Thr Gly 20 25 30 Arg Glu Glu Phe Leu Thr Ala Phe Leu Gln Asn Tyr Gln Leu Ala Tyr 35 40 45 Ser Lys Ala Tyr Pro Arg Leu Leu Ile Ser Ser Leu Ser Glu Ser Pro 50 55 60 Ala Ser Val Ser Ile Leu Ser Gln Ala Asp Asn Thr Ser Lys Lys Val 65 70 75 80 Thr Val Arg Pro Gly Glu Ser Val Met Val Asn Ile Ser Ala Lys Ala 85 90 95 Glu Met Ile Gly Ser Lys Ile Phe Gln His Ala Val Val Ile His Ser 100 105 110 Asp Tyr Ala Ile Ser Val Gln Ala Leu Asn Ala Lys Pro Asp Thr Ala 115 120 125 Glu Leu Thr Leu Leu Arg Pro Ile Gln Ala Leu Gly Thr Glu Tyr Phe 130 135 140 Val Leu Thr Pro Pro Gly Thr Ser Ala Arg Asn Val Lys Glu Phe Ala 145 150 155 160 Val Val Ala Gly Ala Ala Gly Ala Ser Val Ser Val Thr Leu Lys Gly 165 170 175 Ser Val Thr Phe Asn Gly Lys Phe Tyr Pro Ala Gly Asp Val Leu Arg 180 185 190 Val Thr Leu Gln Pro Tyr Asn Val Ala Gln Leu Gln Ser Ser Val Asp 195 200 205 Leu Ser Gly Ser Lys Val Thr Ala Ser Ser Pro Val Ala Val Leu Ser 210 215 220 Gly His Ser Cys Ala Gln Lys His Thr Thr Cys Asn His Val Val Glu 225 230 235 240 Gln Leu Leu Pro Thr Ser Ala Trp Gly Thr His Tyr Val Val Pro Thr 245 250 255 Leu Ala Ser Gln Ser Arg Tyr Asp Leu Ala Phe Val Val Ala Ser Gln 260 265 270 Ala Thr Lys Leu Thr Tyr Asn His Gly Gly Ile Thr Gly Ser Arg Gly 275 280 285 Leu Gln Ala Gly Asp Val Val Glu Phe Glu Val Arg Pro Ser Trp Pro 290 295 300 Leu Tyr Leu Ser Ala Asn Val Gly Ile Gln Val Leu Leu Phe Gly Thr 305 310 315 320 Gly Ala Ile Arg Asn Glu Val Thr Tyr Asp Pro Tyr Leu Val Leu Ile 325 330 335 Pro Asp Val Ala Ala Tyr Cys Pro Ala Tyr Val Val Lys Ser Val Pro 340 345 350 Gly Cys Glu Gly Val Ala Leu Val Val Ala Gln Thr Lys Ala Ile Ser 355 360 365 Gly Leu Thr Ile Asp Gly His Ala Val Gly Ala Lys Leu Thr Trp Glu 370 375 380 Ala Val Pro Gly Ser Glu Phe Ser Tyr Ala Glu Val Glu Leu Gly Thr 385 390 395 400 Ala Asp Met Ile His Thr Ala Glu Ala Thr Thr Asn Leu Gly Leu Leu 405 410 415 Thr Phe Gly Leu Ala Lys Ala Ile Gly Tyr Ala Thr Ala Ala Asp Cys 420 425 430 Gly Arg Thr Val Leu Ser Pro Val Glu Pro Ser Cys Glu Gly Met Gln 435 440 445 Cys Ala Ala Gly Gln Arg Cys Gln Val Val Gly Gly Lys Ala Gly Cys 450 455 460 Val Ala Glu Ser Thr Ala Val Cys Arg Ala Gln Gly Asp Pro His Tyr 465 470 475 480 Thr Thr Phe Asp Gly Arg Arg Tyr Asp Met Met Gly Thr Cys Ser Tyr 485 490 495 Thr Met Val Glu Leu Cys Ser Glu Asp Asp Thr Leu Pro Ala Phe Ser 500 505 510 Val Glu Ala Lys Asn Glu His Arg Gly Ser Arg Arg Val Ser Tyr Val 515 520 525 Gly Leu Val Thr Val Arg Ala Tyr Ser His Ser Val Ser Leu Thr Arg 530 535 540 Gly Glu Val Gly Phe Val Leu Val Asp Asn Gln Arg Ser Arg Leu Pro 545 550 555 560 Val Ser Leu Ser Glu Gly Arg Leu Arg Val Tyr Gln Ser Gly Pro Arg 565 570 575 Ala Val Val Glu Leu Val Phe Gly Leu Val Val Thr Tyr Asp Trp Asp 580 585 590 Cys Gln Leu Ala Leu Ser Leu Pro Ala Arg Phe Gln Asp Gln Val Cys 595 600 605 Gly Leu Cys Gly Asn Tyr Asn Gly Asp Pro Ala Asp Asp Phe Leu Thr 610 615 620 Pro Asp Gly Ala Leu Ala Pro Asp Ala Val Glu Phe Ala Ser Ser Trp 625 630 635 640 Lys Leu Asp Asp Gly Asp Tyr Leu Cys Glu Asp Gly Cys Gln Asn Asn 645 650 655 Cys Pro Ala Cys Thr Pro Gly Gln Ala Gln His Tyr Glu Gly Asp Arg 660 665 670 Leu Cys Gly Met Leu Thr Lys Leu Asp Gly Pro Phe Ala Val Cys His 675 680 685 Asp Thr Leu Asp Pro Arg Pro Phe Leu Glu Gln Cys Val Tyr Asp Leu 690 695 700 Cys Val Val Gly Gly Glu Arg Leu Ser Leu Cys Arg Gly Leu Ser Ala 705 710 715 720 Tyr Ala Gln Ala Cys Leu Glu Leu Gly Ile Ser Val Gly Asp Trp Arg 725 730 735 Ser Pro Ala Asn Cys Pro Leu Ser Cys Pro Ala Asn Ser Arg Tyr Glu 740 745 750 Leu Cys Gly Pro Ala Cys Pro Thr Ser Cys Asn Gly Ala Ala Ala Pro 755 760 765 Ser Asn Cys Ser Gly Arg Pro Cys Val Glu Gly Cys Val Cys Leu Pro 770 775 780 Gly Phe Val Ala Ser Gly Gly Ala Cys Val Pro Ala Ser Ser Cys Gly 785 790 795 800 Cys Thr Phe Gln Gly Leu Gln Leu Ala Pro Gly Gln Glu Val Trp Ala 805 810 815 Asp Glu Leu Cys Gln Arg Arg Cys Thr Cys Asn Gly Ala Thr His Gln 820 825 830 Val Thr Cys Arg Asp Lys Gln Ser Cys Pro Ala Gly Glu Arg Cys Ser 835 840 845 Val Gln Asn Gly Leu Leu Gly Cys Tyr Pro Asp Arg Phe Gly Thr Cys 850 855 860 Gln Gly Ser Gly Asp Pro His Tyr Val Ser Phe Asp Gly Arg Arg Phe 865 870 875 880 Asp Phe Met Gly Thr Cys Thr Tyr Leu Leu Val Gly Ser Cys Gly Gln 885 890 895 Asn Ala Ala Leu Pro Ala Phe Arg Val Leu Val Glu Asn Glu His Arg 900 905 910 Gly Ser Gln Thr Val Ser Tyr Thr Arg Ala Val Arg Val Glu Ala Arg 915 920 925 Gly Val Lys Val Ala Val Arg Arg Glu Tyr Pro Gly Gln Val Leu Val 930 935 940 Asp Asp Val Leu Gln Tyr Leu Pro Phe Gln Ala Ala Asp Gly Gln Val 945 950 955 960 Gln Val Phe Arg Gln Gly Arg Asp Ala Val Val Arg Thr Asp Phe Gly 965 970 975 Leu Thr Val Thr Tyr Asp Trp Asn Ala Arg Val Thr Ala Lys Val Pro 980 985 990 Ser Ser Tyr Ala Glu Ala Leu Cys Gly Leu Cys Gly Asn Phe Asn Gly 995 1000 1005 Asp Pro Ala Asp Asp Leu Ala Leu Arg Gly Gly Gly Gln Ala Ala Asn 1010 1015 1020 Ala Leu Ala Phe Gly Asn Ser Trp Gln Glu Glu Thr Arg Pro Gly Cys 1025 1030 1035 1040 Gly Ala Thr Glu Pro Gly Asp Cys Pro Lys Leu Asp Ser Leu Val Ala 1045 1050 1055 Gln Gln Leu Gln Ser Lys Asn Glu Cys Gly Ile Leu Ala Asp Pro Lys 1060 1065 1070 Gly Pro Phe Arg Glu Cys His Ser Lys Leu Asp Pro Gln Gly Ala Val 1075 1080 1085 Arg Asp Cys Val Tyr Asp Arg Cys Leu Leu Pro Gly Gln Ser Gly Pro 1090 1095 1100 Leu Cys Asp Ala Leu Ala Thr Tyr Ala Ala Ala Cys Gln Ala Ala Gly 1105 1110 1115 1120 Ala Thr Val His Pro Trp Arg Ser Glu Glu Leu Cys Pro Leu Ser Cys 1125 1130 1135 Pro Pro His Ser His Tyr Glu Ala Cys Ser Tyr Gly Cys Pro Leu Ser 1140 1145 1150 Cys Gly Asp Leu Pro Val Pro Gly Gly Cys Gly Ser Glu Cys His Glu 1155 1160 1165 Gly Cys Val Cys Asp Glu Gly Phe Ala Leu Ser Gly Glu Ser Cys Leu 1170 1175 1180 Pro Leu Ala Ser Cys Gly Cys Val His Gln Gly Thr Tyr His Pro Pro 1185 1190 1195 1200 Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys Asp Ser Leu Cys His Cys 1205 1210 1215 Gln Glu Gly Gly Leu Val Ser Cys Glu Ser Ser Ser Cys Gly Pro His 1220 1225 1230 Glu Ala Cys Gln Pro Ser Gly Gly Ser Leu Gly Cys Val Ala Val Gly 1235 1240 1245 Ser Ser Thr Cys Gln Ala Ser Gly Asp Pro His Tyr Thr Thr Phe Asp 1250 1255 1260 Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Val Tyr Val Leu Ala Gln 1265 1270 1275 1280 Thr Cys Gly Thr Arg Pro Gly Leu His Arg Phe Ala Val Leu Gln Glu 1285 1290 1295 Asn Val Ala Trp Gly Asn Gly Arg Val Ser Val Thr Arg Val Ile Thr 1300 1305 1310 Val Gln Val Ala Asn Phe Thr Leu Arg Leu Glu Gln Arg Gln Trp Lys 1315 1320 1325 Val Thr Val Asn Gly Val Asp Met Lys Leu Pro Val Val Leu Ala Asn 1330 1335 1340 Gly Gln Ile Arg Ala Ser Gln His Gly Ser Asp Val Val Ile Glu Thr 1345 1350 1355 1360 Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu Val Tyr Tyr Val Arg Val 1365 1370 1375 Thr Val Pro Gly Asn Tyr Tyr Gln Gln Met Cys Gly Leu Cys Gly Asn 1380 1385 1390 Tyr Asn Gly Asp Pro Lys Asp Asp Phe Gln Lys Pro Asn Gly Ser Gln 1395 1400 1405 Ala Gly Asn Ala Asn Glu Phe Gly Asn Ser Trp Glu Glu Val Val Pro 1410 1415 1420 Asp Ser Pro Cys Leu Pro Pro Thr Pro Cys Pro Pro Gly Ser Glu Asp 1425 1430 1435 1440 Cys Ile Pro Ser His Lys Cys Pro Pro Glu Leu Glu Lys Lys Tyr Gln 1445 1450 1455 Lys Glu Glu Phe Cys Gly Leu Leu Ser Ser Pro Thr Gly Pro Leu Ser 1460 1465 1470 Ser Cys His Lys Leu Val Asp Pro Gln Gly Pro Leu Lys Asp Cys Ile 1475 1480 1485 Phe Asp Leu Cys Leu Gly Gly Gly Asn Leu Ser Ile Leu Cys Ser Asn 1490 1495 1500 Ile His Ala Tyr Val Ser Ala Cys Gln Ala Ala Gly Gly His Val Glu 1505 1510 1515 1520 Pro Trp Arg Thr Glu Thr Phe Cys Pro Met Glu Cys Pro Pro Asn Ser 1525 1530 1535 His Tyr Glu Leu Cys Ala Asp Thr Cys Ser Leu Gly Cys Ser Ala Leu 1540 1545 1550 Ser Ala Pro Pro Gln Cys Gln Asp Gly Cys Ala Glu Gly Cys Gln Cys 1555 1560 1565 Asp Ser Gly Phe Leu Tyr Asn Gly Gln Ala Cys Val Pro Ile Gln Gln 1570 1575 1580 Cys Gly Cys Tyr His Asn Gly Val Tyr Tyr Glu Pro Glu Gln Thr Val 1585 1590 1595 1600 Leu Ile Asp Asn Cys Arg Gln Gln Cys Thr Cys His Ala Gly Lys Gly 1605 1610 1615 Met Val Cys Gln Glu His Ser Cys Lys Pro Gly Gln Val Cys Gln Pro 1620 1625 1630 Ser Gly Gly Ile Leu Ser Cys Val Thr Lys Asp Pro Cys His Gly Val 1635 1640 1645 Thr Cys Arg Pro Gln Glu Thr Cys Lys Glu Gln Gly Gly Gln Gly Val 1650 1655 1660 Cys Leu Pro Asn Tyr Glu Ala Thr Cys Trp Leu Trp Gly Asp Pro His 1665 1670 1675 1680 Tyr His Ser Phe Asp Gly Arg Lys Phe Asp Phe Gln Gly Thr Cys Asn 1685 1690 1695 Tyr Val Leu Ala Thr Thr Gly Cys Pro Gly Val Ser Thr Gln Gly Leu 1700 1705 1710 Thr Pro Phe Thr Val Thr Thr Lys Asn Gln Asn Arg Gly Asn Pro Ala 1715 1720 1725 Val Ser Tyr Val Arg Val Val Thr Val Ala Ala Leu Gly Thr Asn Ile 1730 1735 1740 Ser Ile His Lys Asp Glu Ile Gly Lys Val Arg Val Asn Gly Val Leu 1745 1750 1755 1760 Thr Ala Leu Pro Val Ser Val Ala Asp Gly Arg Ile Ser Val Thr Gln 1765 1770 1775 Gly Ala Ser Lys Ala Leu Leu Val Ala Asp Phe Gly Leu Gln Val Ser 1780 1785 1790 Tyr Asp Trp Asn Trp Arg Val Asp Val Thr Leu Pro Ser Ser Tyr His 1795 1800 1805 Gly Ala Val Cys Gly Leu Cys Gly Asn Met Asp Arg Asn Pro Asn Asn 1810 1815 1820 Asp Gln Val Phe Pro Asn Gly Thr Leu Ala Pro Ser Ile Pro Ile Trp 1825 1830 1835 1840 Gly Gly Ser Trp Arg Ala Pro Gly Trp Asp Pro Leu Cys Trp Asp Glu 1845 1850 1855 Cys Arg Gly Ser Cys Pro Thr Cys Pro Glu Asp Arg Leu Glu Gln Tyr 1860 1865 1870 Glu Gly Pro Gly Phe Cys Gly Pro Leu Ala Pro Gly Thr Gly Gly Pro 1875 1880 1885 Phe Thr Thr Cys His Ala His Val Pro Pro Glu Ser Phe Phe Lys Gly 1890 1895 1900 Cys Val Leu Asp Val Cys Met Gly Gly Gly Asp Arg Asp Ile Leu Cys 1905 1910 1915 1920 Lys Ala Leu Ala Ser Tyr Val Ala Ala Cys Gln Ala Ala Gly Val Val 1925 1930 1935 Ile Glu Asp Trp Arg Ala Gln Val Gly Cys Glu Ile Thr Cys Pro Glu 1940 1945 1950 Asn Ser His Tyr Glu Val Cys Gly Pro Pro Cys Pro Ala Ser Cys Pro 1955 1960 1965 Ser Pro Ala Pro Leu Thr Thr Pro Ala Val Cys Glu Gly Pro Cys Val 1970 1975 1980 Glu Gly Cys Gln Cys Asp Ala Gly Phe Val Leu Ser Ala Asp Arg Cys 1985 1990 1995 2000 Val Pro Leu Asn Asn Gly Cys Gly Cys Trp Ala Asn Gly Thr Tyr His 2005 2010 2015 Glu Ala Gly Ser Glu Phe Trp Ala Asp Gly Thr Cys Ser Gln Trp Cys 2020 2025 2030 Arg Cys Gly Pro Gly Gly Gly Ser Leu Val Cys Thr Pro Ala Ser Cys 2035 2040 2045 Gly Leu Gly Glu Val Cys Gly Leu Leu Pro Ser Gly Gln His Gly Cys 2050 2055 2060 Gln Pro Val Ser Thr Ala Glu Cys Gln Ala Trp Gly Asp Pro His Tyr 2065 2070 2075 2080 Val Thr Leu Asp Gly His Arg Phe Asn Phe Gln Gly Thr Cys Glu Tyr 2085 2090 2095 Leu Leu Ser Ala Pro Cys His Gly Pro Pro Leu Gly Ala Glu Asn Phe 2100 2105 2110 Thr Val Thr Val Ala Asn Glu His Arg Gly Ser Gln Ala Val Ser Tyr 2115 2120 2125 Thr Arg Ser Val Thr Leu Gln Ile Tyr Asn His Ser Leu Thr Leu Ser 2130 2135 2140 Ala Arg Trp Pro Arg Lys Leu Gln Val Asp Gly Val Phe Val Thr Leu 2145 2150 2155 2160 Pro Phe Gln Leu Asp Ser Leu Leu His Ala His Leu Ser Gly Ala Asp 2165 2170 2175 Val Val Val Thr Thr Thr Ser Gly Leu Ser Leu Ala Phe Asp Gly Asp 2180 2185 2190 Ser Phe Val Arg Leu Arg Val Pro Ala Ala Tyr Ala Gly Ser Leu Cys 2195 2200 2205 Gly Leu Cys Gly Asn Tyr Asn Gln Asp Pro Ala Asp Asp Leu Lys Ala 2210 2215 2220 Val Gly Gly Lys Pro Ala Gly Trp Gln Val Gly Gly Ala Gln Gly Cys 2225 2230 2235 2240 Gly Glu Cys Val Ser Lys Pro Cys Pro Ser Pro Cys Thr Pro Glu Gln 2245 2250 2255 Gln Glu Ser Phe Gly Gly Pro Asp Ala Cys Gly Val Ile Ser Ala Thr 2260 2265 2270 Asp Gly Pro Leu Ala Pro Cys His Gly Leu Val Pro Pro Ala Gln Tyr 2275 2280 2285 Phe Gln Gly Cys Leu Leu Asp Ala Cys Gln Val Gln Gly His Pro Gly 2290 2295 2300 Gly Leu Cys Pro Ala Val Ala Thr Tyr Val Ala Ala Cys Gln Ala Ala 2305 2310 2315 2320 Gly Ala Gln Leu Arg Glu Trp Arg Arg Pro Asp Phe Cys Pro Phe Gln 2325 2330 2335 Cys Pro Ala His Ser His Tyr Glu Leu Cys Gly Asp Ser Cys Pro Gly 2340 2345 2350 Ser Cys Pro Ser Leu Ser Ala Pro Glu Gly Cys Glu Ser Ala Cys Arg 2355 2360 2365 Glu Gly Cys Val Cys Asp Ala Gly Phe Val Leu Ser Gly Asp Thr Cys 2370 2375 2380 Val Pro Val Gly Gln Cys Gly Cys Leu His Asp Asp Arg Tyr Tyr Pro 2385 2390 2395 2400 Leu Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys Asp Ser Leu Cys Arg 2405 2410 2415 Cys Arg Glu Gly Gly Glu Val Ser Cys Glu Pro Ser Ser Cys Gly Pro 2420 2425 2430 His Glu Thr Cys Arg Pro Ser Gly Gly Ser Leu Gly Cys Val Ala Val 2435 2440 2445 Gly Ser Thr Thr Cys Gln Ala Ser Gly Asp Pro His Tyr Thr Thr Phe 2450 2455 2460 Asp Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Val Tyr Val Leu Ala 2465 2470 2475 2480 Gln Thr Cys Gly Thr Arg Pro Gly Leu His Arg Phe Ala Val Leu Gln 2485 2490 2495 Glu Asn Val Ala Trp Gly Asn Gly Arg Val Ser Val Thr Arg Val Ile 2500 2505 2510 Thr Val Gln Val Ala Asn Phe Thr Leu Arg Leu Glu Gln Arg Gln Trp 2515 2520 2525 Lys Val Thr Val Asn Gly Val Asp Met Lys Leu Pro Val Val Leu Ala 2530 2535 2540 Asn Gly Gln Ile Arg Ala Ser Gln His Gly Ser Asp Val Val Ile Glu 2545 2550 2555 2560 Thr Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu Val Tyr Tyr Val Arg 2565 2570 2575 Val Thr Val Pro Gly Asn Tyr Tyr Gln Leu Met Cys Gly Leu Cys Gly 2580 2585 2590 Asn Tyr Asn Gly Asp Pro Lys Asp Asp Phe Gln Lys Pro Asn Gly Ser 2595 2600 2605 Gln Ala Gly Asn Ala Asn Glu Phe Gly Asn Ser Trp Glu Glu Val Val 2610 2615 2620 Pro Asp Ser Pro Cys Leu Pro Pro Pro Thr Cys Pro Pro Gly Ser Glu 2625 2630 2635 2640 Gly Cys Ile Pro Ser Glu Glu Cys Pro Pro Glu Leu Glu Lys Lys Tyr 2645 2650 2655 Gln Lys Glu Glu Phe Cys Gly Leu Leu Ser Ser Pro Thr Gly Pro Leu 2660 2665 2670 Ser Ser Cys His Lys Leu Val Asp Pro Gln Gly Pro Leu Lys Asp Cys 2675 2680 2685 Ile Phe Asp Leu Cys Leu Gly Gly Gly Asn Leu Ser Ile Leu Cys Ser 2690 2695 2700 Asn Ile His Ala Tyr Val Ser Ala Cys Gln Ala Ala Gly Gly His Val 2705 2710 2715 2720 Glu Pro Trp Arg Asn Glu Thr Phe Cys Pro Met Glu Cys Pro Gln Asn 2725 2730 2735 Ser His Tyr Glu Leu Cys Ala Asp Thr Cys Ser Leu Gly Cys Ser Ala 2740 2745 2750 Leu Ser Ala Pro Leu Gln Cys Pro Asp Gly Cys Ala Glu Gly Cys Gln 2755 2760 2765 Cys Asp Ser Gly Phe Leu Tyr Asn Gly Gln Ala Cys Val Pro Ile Gln 2770 2775 2780 Gln Cys Gly Cys Tyr His Asn Gly Ala Tyr Tyr Glu Pro Glu Gln Thr 2785 2790 2795 2800 Val Leu Ile Asp Asn Cys Arg Gln Gln Cys Thr Cys His Ala Gly Lys 2805 2810 2815 Val Val Val Cys Gln Glu His Ser Cys Lys Pro Gly Gln Val Cys Gln 2820 2825 2830 Pro Ser Gly Gly Ile Leu Ser Cys Val Thr Lys Asp Pro Cys His Gly 2835 2840 2845 Val Thr Cys Arg Pro Gln Glu Thr Cys Lys Glu Gln Gly Gly Gln Gly 2850 2855 2860 Val Cys Leu Pro Asn Tyr Glu Ala Thr Cys Trp Leu Trp Gly Asp Pro 2865 2870 2875 2880 His Tyr His Ser Phe Asp Gly Arg Lys Phe Asp Phe Gln Gly Thr Cys 2885 2890 2895 Asn Tyr Val Leu Ala Thr Thr Gly Cys Pro Gly Val Ser Thr Gln Gly 2900 2905 2910 Leu Thr Pro Phe Thr Val Thr Thr Lys Asn Gln Asn Arg Gly Asn Pro 2915 2920 2925 Ala Val Ser Tyr Val Arg Val Val Thr Val Ala Ala Leu Gly Thr Asn 2930 2935 2940 Ile Ser Ile His Lys Asp Glu Ile Gly Lys Val Arg Val Asn Gly Val 2945 2950 2955 2960 Leu Thr Ala Leu Pro Val Ser Val Ala Asp Gly Arg Ile Ser Val Ala 2965 2970 2975 Gln Gly Ala Ser Lys Ala Leu Leu Val Ala Asp Phe Gly Leu Gln Val 2980 2985 2990 Ser Tyr Asp Trp Asn Trp Arg Val Asp Val Thr Leu Pro Ser Ser Tyr 2995 3000 3005 His Gly Ala Val Cys Gly Leu Cys Gly Asn Met Asp Arg Asn Pro Asn 3010 3015 3020 Asn Asp Gln Val Phe Pro Asn Gly Thr Leu Ala Pro Ser Ile Pro Ile 3025 3030 3035 3040 Trp Gly Gly Ser Trp Arg Ala Pro Gly Trp Asp Pro Leu Cys Trp Asp 3045 3050 3055 Glu Cys Arg Gly Ser Cys Pro Thr Cys Pro Glu Asp Arg Leu Glu Gln 3060 3065 3070 Tyr Glu Gly Pro Gly Phe Cys Gly Pro Leu Ala Pro Gly Thr Gly Gly 3075 3080 3085 Pro Phe Thr Thr Cys His Ala His Val Pro Pro Glu Ser Phe Phe Lys 3090 3095 3100 Gly Cys Val Leu Asp Val Cys Met Gly Gly Gly Asp His Asp Ile Leu 3105 3110 3115 3120 Cys Lys Ala Leu Ala Ser Tyr Val Ala Ala Cys Gln Ala Ala Gly Val 3125 3130 3135 Val Ile Glu Asp Trp Arg Ala Gln Val Gly Cys Glu Ile Thr Cys Pro 3140 3145 3150 Glu Asn Ser His Tyr Glu Val Cys Gly Pro Pro Cys Pro Ala Ser Cys 3155 3160 3165 Pro Ser Pro Ala Pro Leu Thr Thr Pro Ala Val Cys Glu Gly Pro Cys 3170 3175 3180 Val Glu Gly Cys Gln Cys Asp Ala Gly Phe Val Leu Ser Ala Asp Arg 3185 3190 3195 3200 Cys Val Pro Leu Asn Asn Gly Cys Gly Cys Trp Ala Asn Gly Thr Tyr 3205 3210 3215 His Glu Ala Gly Ser Glu Phe Trp Ala Asp Gly Thr Cys Ser Gln Trp 3220 3225 3230 Cys Arg Cys Gly Pro Gly Gly Gly Ser Leu Val Cys Thr Pro Ala Ser 3235 3240 3245 Cys Gly Leu Gly Glu Val Cys Gly Leu Leu Pro Ser Gly Gln His Gly 3250 3255 3260 Cys Gln Pro Val Ser Thr Ala Glu Cys Gln Ala Trp Gly Asp Pro His 3265 3270 3275 3280 Tyr Val Thr Leu Asp Gly His Arg Phe Asp Phe Gln Gly Thr Cys Glu 3285 3290 3295 Tyr Leu Leu Ser Ala Pro Cys His Gly Pro Pro Leu Gly Ala Glu Asn 3300 3305 3310 Phe Thr Val Thr Val Ala Asn Glu His Arg Gly Ser Gln Ala Val Ser 3315 3320 3325 Tyr Thr Arg Ser Val Thr Leu Gln Ile Tyr Asn His Ser Leu Thr Leu 3330 3335 3340 Ser Ala Arg Trp Pro Arg Lys Leu Gln Val Asp Gly Val Phe Val Thr 3345 3350 3355 3360 Leu Pro Phe Gln Leu Asp Ser Leu Leu His Ala His Leu Ser Gly Ala 3365 3370 3375 Asp Val Val Val Thr Thr Thr Ser Gly Leu Ser Leu Ala Phe Asp Gly 3380 3385 3390 Asp Ser Phe Val Arg Leu Arg Val Pro Ala Ala Tyr Ala Gly Ser Leu 3395 3400 3405 Cys Gly Leu Cys Gly Asn Tyr Asn Gln Asp Pro Ala Asp Asp Leu Lys 3410 3415 3420 Ala Val Gly Gly Lys Pro Ala Gly Trp Gln Val Gly Gly Ala Gln Gly 3425 3430 3435 3440 Cys Gly Glu Cys Val Ser Lys Pro Cys Pro Ser Pro Cys Thr Pro Glu 3445 3450 3455 Gln Gln Glu Ser Phe Gly Gly Pro Asp Ala Cys Gly Val Ile Ser Ala 3460 3465 3470 Thr Asp Gly Pro Leu Ala Pro Cys His Gly Leu Val Pro Pro Ala Gln 3475 3480 3485 Tyr Phe Gln Gly Cys Leu Leu Asp Ala Cys Gln Val Gln Gly His Pro 3490 3495 3500 Gly Gly Leu Cys Pro Ala Val Ala Thr Tyr Val Ala Ala Cys Gln Ala 3505 3510 3515 3520 Ala Gly Ala Gln Leu Arg Glu Trp Arg Arg Pro Asp Phe Cys Pro Phe 3525 3530 3535 Gln Cys Pro Ala His Ser His Tyr Glu Leu Cys Gly Asp Ser Cys Pro 3540 3545 3550 Gly Ser Cys Pro Ser Leu Ser Ala Pro Glu Gly Cys Glu Ser Ala Cys 3555 3560 3565 Arg Glu Gly Cys Val Cys Asp Ala Gly Phe Val Leu Ser Gly Asp Thr 3570 3575 3580 Cys Val Pro Val Gly Gln Cys Gly Cys Leu His Asp Asp Arg Tyr Tyr 3585 3590 3595 3600 Pro Leu Gly Gln Thr Phe Tyr Pro Gly Pro Gly Cys Asp Ser Leu Cys 3605 3610 3615 Arg Cys Arg Glu Gly Gly Glu Val Ser Cys Glu Pro Ser Ser Cys Gly 3620 3625 3630 Pro His Glu Thr Cys Arg Pro Ser Gly Gly Ser Leu Gly Cys Val Ala 3635 3640 3645 Val Gly Ser Thr Thr Cys Gln Ala Ser Gly Asp Pro His Tyr Thr Thr 3650 3655 3660 Phe Asp Gly Arg Arg Phe Asp Phe Met Gly Thr Cys Val Tyr Val Leu 3665 3670 3675 3680 Ala Gln Thr Cys Gly Thr Arg Pro Gly Leu His Arg Phe Ala Val Leu 3685 3690 3695 Gln Glu Asn Val Ala Trp Gly Asn Gly Arg Val Ser Val Thr Arg Val 3700 3705 3710 Ile Thr Val Gln Val Ala Asn Phe Thr Leu Arg Leu Glu Gln Arg Gln 3715 3720 3725 Trp Lys Val Thr Val Asn Gly Val Asp Met Lys Leu Pro Val Val Leu 3730 3735 3740 Ala Asn Gly Gln Ile Arg Ala Ser Gln His Gly Ser Asp Val Val Ile 3745 3750 3755 3760 Glu Thr Asp Phe Gly Leu Arg Val Ala Tyr Asp Leu Val Tyr Tyr Val 3765 3770 3775 Arg Val Thr Val Pro Gly Asn Tyr Tyr Gln Leu Met Cys Gly Leu Cys 3780 3785 3790 Gly Asn Tyr Asn Gly Asp Pro Lys Asp Asp Phe Gln Lys Pro Asn Gly 3795 3800 3805 Ser Gln Ala Gly Asn Ala Asn Glu Phe Gly Asn Ser Trp Glu Glu Val 3810 3815 3820 Val Pro Asp Ser Pro Cys Leu Pro Pro Pro Thr Cys Pro Pro Gly Ser 3825 3830 3835 3840 Glu Gly Cys Ile Pro Ser Glu Glu Cys Pro Pro Glu Leu Glu Lys Lys 3845 3850 3855 Tyr Gln Lys Glu Glu Phe Cys Gly Leu Leu Ser Ser Pro Thr Gly Pro 3860 3865 3870 Leu Ser Ser Cys His Lys Leu Val Asp Pro Gln Gly Pro Leu Lys Asp 3875 3880 3885 Cys Ile Phe Asp Leu Cys Leu Gly Gly Gly Asn Leu Ser Ile Leu Cys 3890 3895 3900 Ser Asn Ile His Ala Tyr Val Ser Ala Cys Gln Ala Ala Gly Gly His 3905 3910 3915 3920 Val Glu Pro Trp Arg Asn Glu Thr Phe Cys Pro Met Glu Cys Pro Gln 3925 3930 3935 Asn Ser His Tyr Glu Leu Cys Ala Asp Thr Cys Ser Leu Gly Cys Ser 3940 3945 3950 Ala Leu Ser Ala Pro Leu Gln Cys Pro Asp Gly Cys Ala Glu Gly Cys 3955 3960 3965 Gln Cys Asp Ser Gly Phe Leu Tyr Asn Gly Gln Ala Cys Val Pro Ile 3970 3975 3980 Gln Gln Cys Gly Cys Tyr His Asn Gly Val Tyr Tyr Glu Pro Glu Gln 3985 3990 3995 4000 Thr Val Leu Ile Asp Asn Cys Arg Gln Gln Cys Thr Cys His Val Gly 4005 4010 4015 Lys Val Val Val Cys Gln Glu His Ser Cys Lys Pro Gly Gln Val Cys 4020 4025 4030 Gln Pro Ser Gly Gly Ile Leu Ser Cys Val Asn Lys Asp Pro Cys His 4035 4040 4045 Gly Val Thr Cys Arg Pro Gln Glu Thr Cys Lys Glu Gln Gly Gly Gln 4050 4055 4060 Gly Val Cys Leu Pro Asn Tyr Glu Ala Thr Cys Trp Leu Trp Gly Asp 4065 4070 4075 4080 Pro His Tyr His Ser Phe Asp Gly Arg Lys Phe Asp Phe Gln Gly Thr 4085 4090 4095 Cys Asn Tyr Val Leu Ala Thr Thr Gly Cys Pro Gly Val Ser Thr Gln 4100 4105 4110 Gly Leu Thr Pro Phe Thr Val Thr Thr Lys Asn Gln Asn Arg Gly Asn 4115 4120 4125 Pro Ala Val Ser Tyr Val Arg Val Val Thr Val Ala Ala Leu Gly Thr 4130 4135 4140 Asn Ile Ser Ile His Lys Asp Glu Ile Gly Lys Val Arg Val Asn Gly 4145 4150 4155 4160 Val Leu Thr Ala Leu Pro Val Ser Val Ala Asp Gly Arg Ile Ser Val 4165 4170 4175 Ala Gln Gly Ala Ser Lys Ala Leu Leu Val Ala Asp Phe Gly Leu Gln 4180 4185 4190 Val Ser Tyr Asp Trp Asn Trp Arg Val Asp Val Thr Leu Pro Ser Ser 4195 4200 4205 Tyr His Gly Ala Val Cys Gly Leu Cys Gly Asn Met Asp Arg Asn Pro 4210 4215 4220 Asn Asn Asp Gln Val Phe Pro Asn Gly Thr Leu Ala Pro Ser Ile Pro 4225 4230 4235 4240 Ile Trp Gly Gly Ser Trp Arg Ala Pro Gly Trp Asp Pro Leu Cys Trp 4245 4250 4255 Asp Glu Cys Arg Gly Ser Cys Pro Thr Cys Pro Glu Asp Arg Leu Glu 4260 4265 4270 Gln Tyr Glu Gly Pro Gly Phe Cys Gly Pro Leu Ala Ser Gly Thr Gly 4275 4280 4285 Gly Pro Phe Thr Thr Cys His Ala His Val Pro Pro Glu Ser Phe Phe 4290 4295 4300 Lys Gly Cys Val Leu Asp Val Cys Met Gly Gly Gly Asp His Asp Ile 4305 4310 4315 4320 Leu Cys Lys Ala Leu Ala Ser Tyr Val Ala Ala Cys Gln Ala Ala Gly 4325 4330 4335 Val Val Ile Glu Asp Trp Arg Ala Gln Val Gly Cys Glu Ile Thr Cys 4340 4345 4350 Pro Glu Asn Ser His Tyr Glu Val Cys Gly Pro Pro Cys Pro Ala Ser 4355 4360 4365 Cys Pro Ser Pro Ala Pro Leu Thr Thr Pro Ala Val Cys Glu Gly Pro 4370 4375 4380 Cys Val Glu Gly Cys Gln Cys Asp Ala Gly Phe Val Leu Ser Ala Asp 4385 4390 4395 4400 Arg Cys Val Pro Leu Asn Asn Gly Cys Gly Cys Trp Ala Asn Gly Thr 4405 4410 4415 Tyr His Glu Ala Gly Ser Glu Phe Trp Ala Asp Gly Thr Cys Ser Gln 4420 4425 4430 Trp Cys Arg Cys Gly Pro Gly Gly Gly Ser Leu Val Cys Thr Pro Ala 4435 4440 4445 Ser Cys Gly Leu Gly Glu Val Cys Gly Leu Leu Pro Ser Gly Gln His 4450 4455 4460 Ser Cys Gln Pro Val Ser Thr Ala Glu Cys Gln Ala Trp Gly Asp Pro 4465 4470 4475 4480 His Tyr Val Thr Leu Asp Gly His Arg Phe Asp Phe Gln Gly Thr Cys 4485 4490 4495 Glu Tyr Leu Leu Ser Ala Pro Cys His Gly Pro Pro Leu Gly Ala Glu 4500 4505 4510 Asn Phe Thr Val Thr Val Ala Asn Glu His Arg Gly Ser Gln Ala Val 4515 4520 4525 Ser Tyr Thr Arg Ser Val Thr Leu Gln Ile Tyr Asn His Ser Leu Thr 4530 4535 4540 Leu Ser Ala Arg Trp Pro Arg Lys Leu Gln Val Asp Gly Val Phe Val 4545 4550 4555 4560 Ala Leu Pro Phe Gln Leu Asp Ser Leu Leu His Ala His Leu Ser Gly 4565 4570 4575 Ala Asp Val Val Val Thr Thr Thr Ser Gly Leu Ser Leu Ala Phe Asp 4580 4585 4590 Gly Asp Ser Phe Val Arg Leu Arg Val Pro Ala Ala Tyr Ala Ala Ser 4595 4600 4605 Leu Cys Gly Leu Cys Gly Asn Tyr Asn Gln Asp Pro Ala Asp Asp Leu 4610 4615 4620 Lys Ala Val Gly Gly Lys Pro Ala Gly Trp Gln Val Gly Gly Ala Gln 4625 4630 4635 4640 Gly Cys Gly Glu Cys Val Ser Lys Pro Cys Pro Ser Pro Cys Thr Pro 4645 4650 4655 Glu Gln Gln Glu Ser Phe Gly Gly Pro Asp Ala Cys Gly Val Ile Ser 4660 4665 4670 Ala Thr Asp Gly Pro Leu Ala Pro Cys His Gly Leu Val Pro Pro Ala 4675 4680 4685 Gln Tyr Phe Gln Gly Cys Leu Leu Asp Ala Cys Gln Val Gln Gly His 4690 4695 4700 Pro Gly Gly Leu Cys Pro Ala Val Ala Thr Tyr Val Ala Ala Cys Gln 4705 4710 4715 4720 Ala Ala Gly Ala Gln Leu Gly Glu Trp Arg Arg Pro Asp Phe Cys Pro 4725 4730 4735 Leu Gln Cys Pro Ala His Ser His Tyr Glu Leu Cys Gly Asp Ser Cys 4740 4745 4750 Pro Val Ser Cys Pro Ser Leu Ser Ala Pro Glu Gly Cys Glu Ser Ala 4755 4760 4765 Cys Arg Glu Gly Cys Val Cys Asp Ala Gly Phe Val Leu Ser Gly Asp 4770 4775 4780 Thr Cys Val Pro Val Gly Gln Cys Gly Cys Leu His Asp Gly Arg Tyr 4785 4790 4795 4800 Tyr Pro Leu Gly Glu Val Phe Tyr Pro Gly Pro Glu Cys Glu Arg Arg 4805 4810 4815 Cys Glu Cys Gly Pro Gly Gly His Val Thr Cys Gln Glu Gly Ala Ala 4820 4825 4830 Cys Gly Pro His Glu Glu Cys Arg Leu Glu Asp Gly Val Gln Ala Cys 4835 4840 4845 His Ala Thr Gly Cys Gly Arg Cys Leu Ala Asn Gly Gly Ile His Tyr 4850 4855 4860 Ile Thr Leu Asp Gly Arg Val Tyr Asp Leu His Gly Ser Cys Ser Tyr 4865 4870 4875 4880 Val Leu Ala Gln Val Cys His Pro Lys Pro Gly Asp Glu Asp Phe Ser 4885 4890 4895 Ile Val Leu Glu Lys Asn Ala Ala Gly His Leu Gln Arg Leu Leu Val 4900 4905 4910 Thr Val Ala Gly Gln Val Val Ser Leu Ala Gln Gly Gln Gln Val Thr 4915 4920 4925 Val Asp Gly Glu Ala Val Ala Leu Pro Val Ala Val Gly Arg Val Arg 4930 4935 4940 Val Thr Ala Glu Gly Arg Asn Met Val Leu Gln Thr Thr Lys Gly Leu 4945 4950 4955 4960 Arg Leu Leu Phe Asp Gly Asp Ala His Leu Leu Met Ser Ile Pro Ser 4965 4970 4975 Pro Phe Arg Gly Arg Leu Cys Gly Leu Cys Gly Asn Phe Asn Gly Asn 4980 4985 4990 Trp Ser Asp Asp Phe Val Leu Pro Asn Gly Ser Ala Ala Ser Ser Val 4995 5000 5005 Glu Thr Phe Gly Ala Ala Trp Arg Val Pro Gly Ser Ser Lys Gly Cys 5010 5015 5020 Gly Glu Gly Cys Gly Pro Gln Gly Cys Pro Val Cys Leu Ala Glu Glu 5025 5030 5035 5040 Thr Ala Pro Tyr Glu Ser Asn Glu Ala Cys Gly Gln Leu Arg Asn Pro 5045 5050 5055 Gln Gly Pro Phe Ala Thr Cys Gln Ala Val Leu Ser Pro Ser Glu Tyr 5060 5065 5070 Phe Arg Gln Cys Val Tyr Asp Leu Cys Ala Gln Lys Gly Asp Lys Ala 5075 5080 5085 Phe Leu Cys Arg Ser Leu Ala Ala Tyr Thr Ala Ala Cys Gln Ala Ala 5090 5095 5100 Gly Val Ala Val Lys Pro Trp Arg Thr Asp Ser Phe Cys Pro Leu His 5105 5110 5115 5120 Cys Pro Ala His Ser His Tyr Ser Ile Cys Thr Arg Thr Cys Gln Gly 5125 5130 5135 Ser Cys Ala Ala Leu Ser Gly Leu Thr Gly Cys Thr Thr Arg Cys Phe 5140 5145 5150 Glu Gly Cys Glu Cys Asp Asp Arg Phe Leu Leu Ser Gln Gly Val Cys 5155 5160 5165 Ile Pro Val Gln Asp Cys Gly Cys Thr His Asn Gly Arg Tyr Leu Pro 5170 5175 5180 Val Asn Ser Ser Leu Leu Thr Ser Asp Cys Ser Glu Arg Cys Ser Cys 5185 5190 5195 5200 Ser Ser Ser Ser Gly Leu Thr Cys Gln Ala Ala Gly Cys Pro Pro Gly 5205 5210 5215 Arg Val Cys Glu Val Lys Ala Glu Ala Arg Asn Cys Trp Ala Thr Arg 5220 5225 5230 Gly Leu Cys Val Leu Ser Val Gly Ala Asn Leu Thr Thr Phe Asp Gly 5235 5240 5245 Ala Arg Gly Ala Thr Thr Ser Pro Gly Val Tyr Glu Leu Ser Ser Arg 5250 5255 5260 Cys Pro Gly Leu Gln Asn Thr Ile Pro Trp Tyr Arg Val Val Ala Glu 5265 5270 5275 5280 Val Gln Ile Cys His Gly Lys Thr Glu Ala Val Gly Gln Val His Ile 5285 5290 5295 Phe Phe Gln Asp Gly Met Val Thr Leu Thr Pro Asn Lys Gly Val Trp 5300 5305 5310 Val Asn Gly Leu Arg Val Asp Leu Pro Ala Glu Lys Leu Ala Ser Val 5315 5320 5325 Ser Val Ser Arg Thr Pro Asp Gly Ser Leu Leu Val Arg Gln Lys Ala 5330 5335 5340 Gly Val Gln Val Trp Leu Gly Ala Asn Gly Lys Val Ala Val Ile Val 5345 5350 5355 5360 Ser Asn Asp His Ala Gly Lys Leu Cys Gly Ala Cys Gly Asn Phe Asp 5365 5370 5375 Gly Asp Gln Thr Asn Asp Trp His Asp Ser Gln Glu Lys Pro Ala Met 5380 5385 5390 Glu Lys Trp Arg Ala Gln Asp Phe Ser Pro Cys Tyr Gly 5395 5400 5405 49 base pairs nucleic acid single linear other nucleic acid /desc = “primer DNA” not provided 10 GCTGATAGTT CTGCAGGAAG GCTGTGAGGA ATTCCTCTCT GCCAGTGTT 49 33 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 11 GCTCCAGCCC AGAGTATCCA CCAGCTCCAT AGG 33 24 base pairs nucleic acid single linear other nucleic acid /desc = “oligonucleotide” not provided 12 CTAGTTAGTT AGTTAGGGTA CCGC 24 24 base pairs nucleic acid single linear other nucleic acid /desc = “oligonucleotide” not provided 13 GGCCGCGGTA CCCTAACTAA CTAA 24 18 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 14 GCCTGCGTGC CCATCCAG 18 19 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 15 CTCATAGTTG GGCAGCGAC 19 18 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 16 TGTTGGGACG AATGTCGG 18 18 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 17 TCACAGCCAA CCTGTGCC 18 18 base pairs nucleic acid single linear other nucleic acid /desc = “oligonucleotide” not provided 18 AGCTTCTGCA GCCATGGG 18 18 base pairs nucleic acid single linear other nucleic acid /desc = “oligonucleotide” not provided 19 AGACGTCGGT ACCCTTAA 18 19 base pairs nucleic acid single linear other nucleic acid /desc = “oligonucleotide” not provided 20 AGCTTCTGCA GCCATCGGG 19 15 base pairs nucleic acid single linear other nucleic acid /desc = “oligonucleotide” not provided 21 AGACGTCGGT ACCCC 15 19 base pairs nucleic acid single linear other nucleic acid /desc = “oligonucleotide” not provided 22 AGCTTCTGCA GCCATGGGA 19 20 base pairs nucleic acid single linear other nucleic acid /desc = “oligonucleotide” not provided 23 AGACGTCGGT ACCCTCTTAA 20 18 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 24 ACCACTCCTT CGATGGCC 18 20 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 25 ACCTGTAACT ATGTGCTGGC 20 19 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 26 TGGTGGTGAC GGTGAAGGG 19 18 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 27 ACAGCAGGGT TGCCCCGG 18 20 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 28 TGGTGCCGAG GGCAGCCACG 20 17 base pairs nucleic acid single linear other nucleic acid /desc = “primer” not provided 29 TGGGTCACTG AAATCCG 

What is claimed is:
 1. An isolated DNA comprising a base sequence encoding the amino acid sequence represented by SEQ ID NO:7 or a base sequence hybridizable therewith in 5×SSPE, 5×Denhardt's solution, 50% formamide, 0.5% SDS, 100 μg/ml salmon sperm DNA, at 42° C. and washed with 0.2×SSC and 0.2% SDS at 65° C., wherein said hybridizing base sequence encodes a polypeptide which binds to the Fc region of IgG.
 2. A DNA as claimed in claim 1 which has been inserted into a plasmid pNV11-ST (FERM BP-4625).
 3. An isolated DNA comprising a base sequence encoding the amino acid sequence represented by SEQ ID NO:9 or a base sequence hybridizable therewith in 5×SSPE, 5×Denhardt's solution, 50% formamide, 0.5% SDS, 100 μg/ml salmon sperm DNA, at 42° C. and washed with 0.2×SSC and 0.2% SDS at 65° C., wherein said hybridizing base sequence encodes a polypeptide which binds to the Fc region of IgG.
 4. A recombinant vector containing the DNA as claimed in claim 1 or
 3. 5. Procaryotic or eucaryotic host cells transformed by a recombinant vector as claimed in claim
 4. 6. A process for producing a recombinant protein which comprises incubating host cells as claimed in claim 5 and separating and purifying the protein thus produced.
 7. A method for identifying a tissue synthesizing mRNA of the IgG Fc region-binding protein by Northern blotting or in situ hybridization with the use of a DNA as claimed in claim 1 or 3 as a probe.
 8. An isolated DNA comprising: a) an H (nucleotides 9-1352 of SEQ ID NO:8) region and one or more regions selected from the group consisting of R1 (nucleotides 1353-2528 of SEQ ID NO:8), R2 (nucleotides 2529-3692 of SEQ ID NO:8), R3 (nucleotides 3693-4955 of SEQ ID NO:8), R4 (nucleotides 4956-6149 of SEQ ID NO:8), R5 (nucleotides 6150-7295 of SEQ ID NO:8), R6 (nucleotides 7296-8558 of SEQ ID NO:8), R7 (nucleotides 8559-9752 of SEQ ID NO:8), R8 (nucleotides 9753-10898 of SEQ ID NO:8), R9 (nucleotides 10899-12161 of SEQ ID NO:8), R10 (nucleotides 12162-13355 of SEQ ID NO:8), R11 (nucleotides 13356-14504 of SEQ ID NO:8) and R12 (nucleotides 14505-15767 of SEQ ID NO:8); or b) an isolated DNA which hybridizes to said DNA of a) in 5×SSPE, 5×Denhardt's solution, 50% formamide, 0.5% SDS, 100 μg/ml salmon sperm DNA, at 42° C. and washed with 0.2×SSC and 0.2% SDS at 65° C., wherein said hybridizing base sequence encodes a polypeptide which binds to the Fc region of IgG.
 9. An isolated DNA sequence encoding a protein of SEQ ID NO:7 or
 9. 10. An isolated DNA comprising: an H (nucleotides 9-1352 of SEQ ID NO:8) region and one or more regions selected from the group consisting of R1 (nucleotides 1353-2528 of SEQ ID NO:8), R2 (nucleotides 2529-3692 of SEQ ID NO:8), R3 (nucleotides 3693-4955 of SEQ ID NO:8), R4 (nucleotides 4956-6149 of SEQ ID NO:8), R5 (nucleotides 6150-7295 of SEQ ID NO:8), R6 (nucleotides 7296-8558 of SEQ ID NO:8), R7 (nucleotides 8559-9752 of SEQ ID NO:8) R8 (nucleotides 9753-10898 of SEQ ID NO:8), R9 (nucleotides 10899-12161 of SEQ ID NO:8), R10 (nucleotides 12162-13355 of SEQ ID No:8), R11 (nucleotides 13356-14504 of SEQ ID NO:8) and R12 (nucleotides 14505-15767 of SEQ ID NO:8) wherein said DNA encodes a protein which binds to the IgG Fc region. 